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Pou5f1

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POU5F1 is a gene that encodes a transcription factor protein. This protein is involved in the regulation of embryonic stem cell pluripotency and self-renewal. POU5F1 plays a critical role in maintaining the undifferentiated state of embryonic stem cells.

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Lab products found in correlation

Gapdh, by Thermo Fisher Scientific (2 mentions) Nanos3, by Thermo Fisher Scientific (2 mentions) Prdm1, by Thermo Fisher Scientific (2 mentions) Tfap2c, by Thermo Fisher Scientific (2 mentions) Prdm14, by Thermo Fisher Scientific (2 mentions) Taqman universal pcr master mix, by Thermo Fisher Scientific (2 mentions) Superscript 2 reverse transcriptase, by Thermo Fisher Scientific (2 mentions) Rlt buffer, by Qiagen (2 mentions) Rneasy micro kit, by Qiagen (2 mentions) Taqman primer, by Thermo Fisher Scientific (2 mentions) Tubb3, by Thermo Fisher Scientific (2 mentions) Axin2, by Thermo Fisher Scientific (1 mentions) Rnaeasy kit, by Qiagen (1 mentions) Nanog, by Thermo Fisher Scientific (1 mentions) Ccnd1, by Thermo Fisher Scientific (1 mentions)

5 protocols using pou5f1

1

Profiling Pluripotency Genes in Germ Cell-Like Cells

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PGCLCs sorted or harvested from membranes were placed in 350 μL of RLT buffer (QIAGEN) and RNA was extracted using RNeasy micro kit (QIAGEN, 74004). cDNA was synthesized using SuperScript® II Reverse Transcriptase (Invitrogen, 18064-014). Real time quantitative PCR was performed using TaqMan® Universal PCR Master Mix (Applied Biosystems, 4304437) and the expression level of genes-of-interest were normalized to the expression of housekeeping gene GAPDH. The TaqMan probes used in this study include: GAPDH (Applied Biosystems, Hs99999905_m1), NANOS3 (Applied Biosystems, Hs00928455_s1), PRDM1 (Applied Biosystems, hs01068508_m1), TFAP2C (Applied Biosystems, Hs00231476_m1), POU5F1 (Applied Biosystems, Hs03005111_g1), SOX2 (Applied Biosystems, Hs01053049_s1), PRDM14 (Applied Biosystems, Hs01119056_m1).
Gell J.J., Zhao J., Chen D., Hunt T.J, & Clark A.T. (2018). PRDM14 is expressed in germ cell tumors with constitutive overexpression altering human germline differentiation and proliferation. Stem cell research, 27, 46-56.
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2

Profiling Pluripotency Genes in Germ Cell-Like Cells

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PGCLCs sorted or harvested from membranes were placed in 350 μL of RLT buffer (QIAGEN) and RNA was extracted using RNeasy micro kit (QIAGEN, 74004). cDNA was synthesized using SuperScript® II Reverse Transcriptase (Invitrogen, 18064-014). Real time quantitative PCR was performed using TaqMan® Universal PCR Master Mix (Applied Biosystems, 4304437) and the expression level of genes-of-interest were normalized to the expression of housekeeping gene GAPDH. The TaqMan probes used in this study include: GAPDH (Applied Biosystems, Hs99999905_m1), NANOS3 (Applied Biosystems, Hs00928455_s1), PRDM1 (Applied Biosystems, hs01068508_m1), TFAP2C (Applied Biosystems, Hs00231476_m1), POU5F1 (Applied Biosystems, Hs03005111_g1), SOX2 (Applied Biosystems, Hs01053049_s1), PRDM14 (Applied Biosystems, Hs01119056_m1).
Gell J.J., Zhao J., Chen D., Hunt T.J, & Clark A.T. (2018). PRDM14 is expressed in germ cell tumors with constitutive overexpression altering human germline differentiation and proliferation. Stem cell research, 27, 46-56.
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3

Stem Cell Characterization by qPCR

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RNA was isolated via QIAcube (serial number: 50946) and RNeasy Mini Kit protocol “Purification of total RNA from 5 × 106 − 1 × 107 animal cells (with QIAshreddder homogenization)” (Firmware Version FIW-50–001-J_FW_MB.hex and program version FIW-50–002-G_PLC_MB.prs or higher) following manufacturer’s instructions. cDNA was synthesized using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosysystems, Cat. # 4368814) according to manufacturer’s instructions. Quantitative PCR with the Taqman™ primers POU5F1 (Thermofisher, Cat. # 4448892), PAX6 (Thermofisher, Cat. # 4448892), NES (Thermofisher, Cat. # 4448892), TUBB3 (Thermofisher, Cat. # 4448892), and OTX2 (Thermofisher, Cat. # 4448892) was carried out by a liquid handling robotic system and run via Quantstudio 12K Flex (serial number: 285880333).
Kuang Y.L., Munoz A., Nalula G., Santostefano K.E., Sanghez V., Sanchez G., Terada N., Mattis A.N., Iacovino M., Iribarren C., Krauss R.M, & Medina M.W. (2019). Evaluation of commonly used ectoderm markers in iPSC trilineage differentiation. Stem cell research, 37, 101434.
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4

Stem Cell Characterization by qPCR

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RNA was isolated via QIAcube (serial number: 50946) and RNeasy Mini Kit protocol “Purification of total RNA from 5 × 106 − 1 × 107 animal cells (with QIAshreddder homogenization)” (Firmware Version FIW-50–001-J_FW_MB.hex and program version FIW-50–002-G_PLC_MB.prs or higher) following manufacturer’s instructions. cDNA was synthesized using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosysystems, Cat. # 4368814) according to manufacturer’s instructions. Quantitative PCR with the Taqman™ primers POU5F1 (Thermofisher, Cat. # 4448892), PAX6 (Thermofisher, Cat. # 4448892), NES (Thermofisher, Cat. # 4448892), TUBB3 (Thermofisher, Cat. # 4448892), and OTX2 (Thermofisher, Cat. # 4448892) was carried out by a liquid handling robotic system and run via Quantstudio 12K Flex (serial number: 285880333).
Kuang Y.L., Munoz A., Nalula G., Santostefano K.E., Sanghez V., Sanchez G., Terada N., Mattis A.N., Iacovino M., Iribarren C., Krauss R.M, & Medina M.W. (2019). Evaluation of commonly used ectoderm markers in iPSC trilineage differentiation. Stem cell research, 37, 101434.
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5

Transcriptional Profiling of Stem Cell Markers

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Cells were plated in 25 cm2-angled flasks at 1 × 10^6 cells of confluence, 48 h after treatments, cells were scraped from flasks to extract RNA; the extraction was performed with RNAeasy kit (QUIAGEN, Hilden Germany) according to manufacturer’s instructions. Next cDNA was obtained by retro-transcription assay using Revert Aid Synthesis Kit (Thermofisher, United States). For RT-PCR assays, taqman probes used for: (Hs00610060_m1) SFRP1; (Hs00277039_m1) CCND1; (Hs03986777_s1) FZD4; (Hs02387400_g1) NANOG; (Hs99999903_m1) ACTB; (Hs00153408_m1) MYC; (Hs04260367_gH) POU5F1; (Hs00610344_m1) AXIN2; (Hs00602736_s1) SOX2; (Hs99999901_s1) 18S, and (Hs01547250_m1) LEF1 genes were purchased from Thermofisher (Massachusetts, United States).
Losada-García A., Salido-Guadarrama I., Cortes-Ramirez S.A., Cruz-Burgos M., Morales-Pacheco M., Vazquez-Santillan K., Rodriguez-Martinez G., González-Ramírez I., Gonzalez-Covarrubias V., Perez-Plascencia C, & Rodríguez-Dorantes M. (2023). SFRP1 induces a stem cell phenotype in prostate cancer cells. Frontiers in Cell and Developmental Biology, 11, 1096923.
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