Pparg

Manufactured by Thermo Fisher Scientific

Sourced in United States

The PPARG product is a laboratory equipment item used for scientific research purposes. It is a highly sensitive and specific tool designed to detect and analyze the expression of the PPARG gene, which is involved in various biological processes. The PPARG product provides researchers with the necessary tools to study the role of this gene in areas such as cell differentiation, metabolism, and inflammation. This factual description outlines the core function of the PPARG product without making any interpretations or extrapolations about its intended use.

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Anti-PPARg (2 citations) TaqMan Gene Expression Assays for PPARG (2 citations) ON-TARGETplus SMARTpool Human PPARG (2 citations)

15 protocols using pparg

Adipocyte mRNA Expression Analysis

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mRNA was purified from adipocytes or adipose tissue according to the manufacturer’s instructions (Qiagen, Germantown, MD) and quantified using 7900HT Fast Real-Time PCR System and TaqMan fluorogenic detection system (Applied Biosystems, Grand Island, NY). The Lep (Assay ID: Mm00434759_m1), Pref1 (Assay ID: Mm00494477_m1), and Pparg (Assay ID: Mm00440945_m2) validated primers were also purchased from Applied Biosystems (Grand Island, NY). Comparative real-time PCR was performed in triplicate, including no-template controls. Expression was calculated using the comparative Ct method normalized to the TATA box binding protein (TBP, Applied Biosystems, Grand Island, NY, Assay ID: Mm00446973_m1).

DiSilvestro D.J., Melgar-Bermudez E., Yasmeen R., Fadda P., Lee L.J., Kalyanasundaram A., Gilor C.L, & Ziouzenkova O. (2016). Leptin Production by Encapsulated Adipocytes Increases Brown Fat, Decreases Resistin, and Improves Glucose Intolerance in Obese Mice. PLoS ONE, 11(4), e0153198.

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Ileum RNA Isolation and qRT-PCR

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Total RNA from ileum was isolated using the RNeasy minikit according to the manufacturer’s instructions (Qiagen, CA, USA). Reverse transcriptase PCR was performed with a high-capacity cDNA reverse transcription kit (Applied Biosystems, CA, USA). TaqMan assay IDs were as follows: Tbp (Mm01277042_m1), Cnr1 (Mm01212171_s1), Cnr2 (Mm02620087_s1), Ppara (Mm00440939_m1), Pparg (Mm00440940_m1), Abhd6 (Mm00481199_m1), Plcb1 (Mm00479987_m1), and Faah (Mm00515684_m1) (Applied Biosystems, CA, USA). All expression data were normalized by the threshold cycle (2^−ΔΔ^CT) method using Tbp as internal control (36 (link)).

Lacroix S., Pechereau F., Leblanc N., Boubertakh B., Houde A., Martin C., Flamand N., Silvestri C., Raymond F., Di Marzo V, & Veilleux A. (2019). Rapid and Concomitant Gut Microbiota and Endocannabinoidome Response to Diet-Induced Obesity in Mice. mSystems, 4(6), e00407-19.

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Profiling Gene Expression in Immune Cells

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RNA was extracted using RNAeasy columns (Qiagen) and complementary DNA was prepared following the manufacturer’s instructions (Applied Biosystems) and used as the template for either real-time PCR or RT2Profiler PCR Array (330231 PAMM-006ZA, Qiagen). All the primers and probes were provided by Applied Biosystems, and were used on a ViiA 7 Real-Time PCR System (Applied Biosystems). Expression was normalized to GAPDH expression. The following primers and probe mixtures were purchased from Applied Biosystems: Entpd1 (Mm00515447_m1), Ahr (Mm00478930_m1), Hif1a (Mm00468872_m1), Stat1 (Mm00439518_m1), Klf4 (Mm00516104_m1), Pparg (Mm01184322_m1), Vegfa (Mm00437306_m1), Arg1 (Mm00475988_m1), Cd274 (Mm00452054_m1), Tgfb1 (Mm0117820_m1), Il10 (Mm00439614_m1), Tnfa (Mm00443258_m1), Il1b (Mm00434228_m1), Il27 (Mm00461162_m1), Mrc1 (Mm01329363_m1), Tdo2 (Mm00451266_m1), Ido1 (Mm01218007_m1), Clec10a (Mm00546125_g1), Socs2 (Mm00850544_g1), Cyp1b1 (Mm00487229_m1), Cyp1a1 (Mm00487218_m1), Ccr2 (Mm00438270_m1), Ccl2 (Mm00441242_m1), Ccl7 (Mn00443113_m1), hsa-miR-29a (002112), hsa-miR-29b (000413), Gapdh (Mm99999915_g1), AHR (Hs00169233_m1), CYP1B (Hs02382916_s1), MRC1 (Hs00267207_m1), KLF4 (Hs00358836_m1) Entpd1 (Hs00969559_m1), LLGL1 (Hs01017181_m1) and GAPDH (Hs02758991_g1).

Takenaka M.C., Gabriely G., Rothhammer V., Mascanfroni I.D., Wheeler M.A., Chao C.C., Gutiérrez-Vázquez C., Kenison J., Tjon E.C., Barroso A., Vandeventer T., de Lima K.A., Rothweiler S., Mayo L., Ghannam S., Zandee S., Healy L., Sherr D., Farez M.F., Prat A., Antel J., Reardon D.A., Zhang H., Robson S.C., Getz G., Weiner H.L, & Quintana F.J. (2019). Control of tumor-associated macrophages and T cells in glioblastoma via AHR and CD39. Nature neuroscience, 22(5), 729-740.

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Exercise-Induced Macrophage Gene Expression

Cited 1 time

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Immediately (0 h) or 48 h after the last session of exercise protocol, mice were euthanized and macrophages were removed from the peritoneal cavity and aortic arch was harvested. Tissues were macerated in liquid nitrogen and homogenized as proposed by RNeasy^® Mini Kit (Qiagen, Hilden, Germany). The expression of genes was determined by real time quantitative reverse transcription polymerase chain reaction (RT-qPCR) as described before by Pinto et al. (2015) (link) according to Livak and Schmittgen (2001) (link). Using primer by Applied Biosystems (Foster City, CA, United States), the following genes were analyzed: Abca1 (Mm00442646_m1), Abcg1 (Mm00437390_m1), Nr1h3 (Mm01329744_g1), Nr1h2 (Mm00437265_g1), Pparg (Mm01184322_m1), Scarb1 (Mm00450234_m1), Olr1 (Mm00454586_m1), Cd36 (Mm01135198_m1), Tnf (Mm00450234_m1), Il6 (Mm00450234_m1), Il10 (Mm00450234_m1), Ccl2 (Mm00441242_m1), Nos3 (Mm004435217_m1) and Cat (Mm00443258_m1). A gene stability assay was performed and according to a ranking order, Actb (Mm00607939_s1) and Gapdh (Mm99999915_g1) were utilized as endogenous control, respectively for macrophages and aortic arch samples.

Pinto P.R., da Silva K.S., Iborra R.T., Okuda L.S., Gomes-Kjerulf D., Ferreira G.S., Machado-Lima A., Rocco D.D., Nakandakare E.R., Machado U.F., Correa-Giannella M.L., Catanozi S, & Passarelli M. (2018). Exercise Training Favorably Modulates Gene and Protein Expression That Regulate Arterial Cholesterol Content in CETP Transgenic Mice. Frontiers in Physiology, 9, 502.

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Multilineage Gene Expression Analysis

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On day 7 and 21, total RNA was isolated using TRIzol™ reagent according to the manufacturer’s protocol. Reverse transcription of 500 ng RNA was performed using the High-Capacity cDNA Reverse Transcription Kit in the presence of RNase inhibitor. Real-time qPCR was performed with 25 ng of cDNA using an ABI 7500 Real-Time PCR Machine, TaqMan™ Universal PCR Master Mix, and the following human FAM labelled TaqMan® gene expression primers: Sox9 (Hs00165814-m1), OCN (Hs01587814-g1), Runx2 (Hs01047973-m1), ALPL (Hs01029144-m1), PPARg (Hs01115513-m1), Leptin (Ha00174877-m1), LEPR (Hs00174497-m1), Sox7 (Hs00846731-s1), ACAN (Hs00153936-m1), and Col10 (Hs00166657-m1) (all purchased from Applied Biosystems). Beta-actin (Hs01060665-g1) was used as endogenous control. Gene expression was analyzed according to the delta-delta Ct method and are presented as fold-changes (2^{−delta-delta Ct}) of treated samples to non-treated samples.

Tauer J.T., Al-Jallad H., Umebayashi M., Bakhsh D., Rauch D., Tran S.D., Rauch F, & Hamdy R. (2022). Characterization and functional analysis of the adipose tissue-derived stromal vascular fraction of pediatric patients with osteogenesis imperfecta. Scientific Reports, 12, 2414.

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Gene Expression Analysis of Bioprinted Tissues

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Gene expression was analyzed using real-time quantitative polymerase chain reaction (RT-qPCR). Adipose drop tissues at days 0, 7, and 14 of differentiation, as well as bioprinted fiber samples at days (before bioprinting) and at days 6 (myoblast fibers), 7 (endothelial fibers), or 14 (adipocyte fibers) were washed in PBS and total RNA extraction was carried out using the PureLink RNA Micro Kit (Invitrogen, Waltham, USA), with the DNAse step, following the manufacturer’s instructions. Samples’ RNA content was quantified with the NanodropTM spectrometer (N1000, Thermo Fisher Scientific, Waltham, USA). For RT-qPCR, the RNA samples were first submitted to reverse transcription into cDNA using iSCRIPT cDNA synthesis kit (Bio-Rad, Hercules, USA), before being amplified using Taqman probes and reagents (Taqman Fast Advanced Mix, Taqman gene expression assays (FAM): MYH2 (Assay ID: Bt03223147_gH), FABP4 (Assay ID: Bt03213820_m1), CD31/PECAM1 (Assay ID: Bt03215106_m1), PPARG (Assay ID: Bt03217547_m1), and PPIA (Assay ID: Bt03224615_g1), Thermo Fisher Scientific, Waltham, USA). The cDNA synthesis and RT-qPCR reactions were conducted using the StepOnePlus Real-Time PCR System (Thermo Fisher Scientific, Waltham, USA) and the gene expression was normalized by PPIA as the housekeeping gene.

Kang D.H., Louis F., Liu H., Shimoda H., Nishiyama Y., Nozawa H., Kakitani M., Takagi D., Kasa D., Nagamori E., Irie S., Kitano S, & Matsusaki M. (2021). Engineered whole cut meat-like tissue by the assembly of cell fibers using tendon-gel integrated bioprinting. Nature Communications, 12, 5059.

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Quantitative Gene Expression Analysis in Adipocytes

Cited 2 times

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Total RNA was extracted from hWAs adipocytes using an RNeasy Plus kit (74136, Qiagen) according to the manufacturer’s instructions. RNA purity and concentration were measured with Nanodrop (Nanodrop, USA). cDNA was then synthesised using SuperScript IV VILO Master Mix (11756500, Thermo Fisher Scientific). The relative gene expressions were detected using the ViiA7 Real-Time PCR system (PE Applied Biosystems, USA) with pre-designed Taqman assays following the manufacturer’s instructions. The following pre-designed gene expression assays were ordered from Thermo Fisher Scientific: PPARGC1A (Hs00173304_m1), PPARG (Hs01115513_m1), FABP4 (Hs01086177_m1), CEBPA (Hs00269972_s1), SLC2A4 (Hs00168966_m1), CEBPB (Hs00942496_s1), TOMM20 (Hs03276810_g1), TFAM (Hs01073348_g1), PPARGC1B (Hs00370186_m1), LIPE (Hs00193510_m1), ABHD5 (Hs01104373_m1), ACACB (Hs01565914_m1), CPT1B (Hs00189258_m1), CS (Hs02574374_s1), ADIPOQ (Hs00977214_m1), SCD (Hs01682761_m1), SREBF1 (Hs02561944_s1), FASN (Hs01005622_m1), PNPLA2 (Hs00386101_m1), LPL (Hs00173425_m1), MT-CO2 (Hs02596865_g1), HPRT1 (Hs99999909_m1), TBP (Hs00427620_m1) and RPL13A (Hs03043885_g1). The geometric means of TBP, HPRT1 and RPL13A housekeeping gene expression [20 (link)–22 (link)] were used to normalise the expression of genes of interest, and, unless indicated otherwise, the ΔΔC_t method was used to analyse the results.

Huang M., Claussnitzer M., Saadat A., Coral D.E., Kalamajski S, & Franks P.W. (2023). Engineered allele substitution at PPARGC1A rs8192678 alters human white adipocyte differentiation, lipogenesis, and PGC-1α content and turnover. Diabetologia, 66(7), 1289-1305.

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Metabolic Profiling via Real-Time PCR

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Real time PCR was performed using TaqMan Fast Advanced Master Mix and Taqman gene expression assay probes for Arginase 1, Cd206, Acadl, Acadm, Cpt1b, Pparg, Ppargc1b, Ogt, Mgea5, Fasn, and Acaca (Thermo Fisher). Actb served as an endorse control gene. Each reaction was done in triplicate using an Applied Biosystems Step One Plus kit.

Hinshaw D.C., Hanna A., Lama-Sherpa T., Metge B., Kammerud S.C., Benavides G.A., Kumar A., Alsheikh H.A., Mota M., Chen D., Ballinger S.W., Rathmell J.C., Ponnazhagan S., Darley-Usmar V., Samant R.S, & Shevde L.A. (2021). Hedgehog signaling regulates metabolism and polarization of mammary tumor-associated macrophages. Cancer research, 81(21), 5425-5437.

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Quantification of Aquaporin and Lipogenic Gene Expression

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Primary HSC and LX2 cells were homogenized in TRIzol reagent (Thermo Fisher Scientific) and RNA was isolated according to the manufacturer´s protocol. Total RNA (1 µg) was transcribed into cDNA using Superscript II and random hexamer primers (Thermo Fisher Scientific). Gene expression of human AQP3 (NM_004925.4), AQP7 (NM_001170.2), AQP9 (NM_001320635.1), AQP10 (XM_011510104.2), FASN (NM_004104.4), SCD1 (NM_005063.4), PNPLA3 (NM_025225.2), p21 (NM_000389.4), PPARg (NM 005037.5) and SREBP1c (NM_001321096.2) (all Thermo Fisher Scientific) was analyzed by quantitative real-time PCR on an ABI Step One Plus cycler using assays-on-demand kits (TaqMan® Gene Expression Assay, Thermo Fisher Scientific). Each reaction was performed in duplicates and the value of the gene of interest was normalized to human ubiquitin C expression. The comparative threshold cycle (CT) method was used to calculate the relative expression^{46 (link)}.

Tardelli M., Bruschi F.V., Claudel T., Moreno-Viedma V., Halilbasic E., Marra F., Herac M., Stulnig T.M, & Trauner M. (2017). AQP3 is regulated by PPARγ and JNK in hepatic stellate cells carrying PNPLA3 I148M. Scientific Reports, 7, 14661.

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Quantitative gene expression analysis

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Total RNA was extracted using TRIzol reagent (Invitrogen). RNA was converted to cDNA using High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). Quantitative PCR reactions were prepared with TaqMan™ Fast Advanced Master Mix (Thermo Fisher Scientific) and performed on ViiA 7 Real-Time PCR System (Thermo Fisher Scientific). Data were normalized to the content of Cyclophilin A (PPIA), as the endogenous control. TaqMan primers obtained from Thermo Fisher Scientific were: LEP (Hs00174877_m1), PPARG (Hs01115513_m1), CEBPA (Hs00269972), UCP1 (Hs01084772), PPIA (Hs04194521_s1), Adipoq (Mm00456425_m1), PPARG (Mm00440940_m1), Cebpa (Mm00514283_s1), Fabp4 (Mm00445878_m1), Ppia (Mm02342430_g1); TaqMan primers obtained from IDT were: FAM13A (HS.PT.58.3237208), ADIPOQ (Hs.PT.58.39189358), and PGC1A (Hs.PT.58.14965839), Fam13a (Mm.PT.56a.7855516).

Fathzadeh M., Li J., Rao A., Cook N., Chennamsetty I., Seldin M., Zhou X., Sangwung P., Gloudemans M.J., Keller M., Attie A., Yang J., Wabitsch M., Carcamo-Orive I., Tada Y., Lusis A.J., Shin M.K., Molony C.M., McLaughlin T., Reaven G., Montgomery S.B., Reilly D., Quertermous T., Ingelsson E, & Knowles J.W. (2020). FAM13A affects body fat distribution and adipocyte function. Nature Communications, 11, 1465.

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