Hydroxychloroquine

Manufactured by Selleck Chemicals

Sourced in United States

Hydroxychloroquine is a chemical compound used in laboratory research. It functions as a pH-modulating agent, capable of altering the acidity or basicity of cellular environments. This property may be utilized in various experimental settings, such as studies involving cellular processes, signaling pathways, or host-pathogen interactions.

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Lab products found in correlation

Gapdh, by Abcam (2 mentions) β actin, by Abcam (2 mentions) Mk 2206, by Selleck Chemicals (2 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) α tubulin, by Santa Cruz Biotechnology (1 mentions) Silencer select pre designed sirna, by Thermo Fisher Scientific (1 mentions) Bkm120, by Selleck Chemicals (1 mentions) β actin, by Santa Cruz Biotechnology (1 mentions) Phospho akt s473, by Santa Cruz Biotechnology (1 mentions) Phospho foxo3a s253, by Cell Signaling Technology (1 mentions) Pan 14 3 3, by Santa Cruz Biotechnology (1 mentions) Foxo3a, by Cell Signaling Technology (1 mentions) Remdesivir, by Selleck Chemicals (1 mentions) Ly294002, by Selleck Chemicals (1 mentions) Compound c, by Selleck Chemicals (1 mentions) Phospho ampk, by Cell Signaling Technology (1 mentions) Horseradish peroxidase hrp conjugated secondary antibody, by Cell Signaling Technology (1 mentions) Trametinib, by Selleck Chemicals (1 mentions) Mek162, by Selleck Chemicals (1 mentions) P akt, by Abcam (1 mentions)

Spelling variants of this product

Hydroxychloroquine (HCQ) (10 citations) Hydroxychloroquine sulfate (6 citations) Hydroxychloroquine sulfate (HCQ) (3 citations)

6 protocols using hydroxychloroquine

Investigating Autophagy and Apoptosis Regulation

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BKM120 (S2247) and hydroxychloroquine (S4430) were purchased from Selleck Chemicals. BV02 (SML0140) was purchased from Sigma-Aldrich. Antibodies against the following proteins were used in this study: phospho-Akt S473 (Santa Cruz Biotechnology, sc-7985-R), Akt (Santa Cruz Biotechnology, sc-8312), α-tubulin (Santa Cruz Biotechnology, sc-8035), pan-14-3-3 (Santa Cruz Biotechnology, sc-629), 14-3-3ζ (Santa Cruz Biotechnology, sc-1019), p27 (Santa Cruz Biotechnology, sc-1641), β-actin (Santa Cruz Biotechnology, sc-47778), cleaved caspase-3 (Cell Signaling Technology, #9661), phospho-FOXO3a S253 (Cell Signaling Technology, #13129), FOXO3a (Cell Signaling Technology, #2497), LC3B (Abcam, ab48394), ATG7 (MBL, PM039), and di-methyl histone H3 (Lys9) (Cell Signaling Technology, #9753). The LC3B-EGFP expression vector (#11546) and the FHRE-luc reporter vector (#1789) were from Addgene (Cambridge, MA). The siRNAs used in the present study included ATG7 siRNA (Ambion, Silencer® Select Pre-designed siRNA, Cat#s20651), 14-3-3ζ siRNA (Invitrogen, Stealth siRNA™, cat#5480996), FOXO3a siRNA (Invitrogen, Stealth siRNA™, Cat#5436311), and negative control siRNA (Bioneer, SN-1022).

Kim H.J., Lee Yoon S., Young C.K., Hwan Y.K., Ju W, & Cheol S.K. (2016). Subcellular localization of FOXO3a as a potential biomarker of response to combined treatment with inhibitors of PI3K and autophagy in PIK3CA-mutant cancer cells. Oncotarget, 8(4), 6608-6622.

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Antiviral Efficacy Testing of Hydroxychloroquine, Remdesivir, and IFNB1 against SARS-CoV-2

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Hydroxychloroquine (Selleck Chemicals Cat. No. S4430) and Remdesivir (Selleck Chemicals Cat. No. S8932) were dissolved in DMSO to a stock concentration of 10 mM. IFNB1 stock of 10ˆ6 units/ml was provided by Dr. Jay Kolls. To test the efficacy of the drugs against SARS-CoV-2 infection/replication in AT2 cells, cultures were suspended in media containing treated with 10 μM of Hydroxychloroquine or Remdesivir, or 100 units/ml of IFNB1 3 hours prior to SARS-CoV-2 infection. Viral infections were performed as described previously. Drugs were maintained in the media for the duration of culture post infection. alveolar cultures without drug treatment in the presence or absence (mock) of viral infections were included as controls.

Mulay A., Konda B., Garcia G J.r., Yao C., Beil S., Villalba J.M., Koziol C., Sen C., Purkayastha A., Kolls J.K., Pociask D.A., Pessina P., de Aja J.S., Garcia-de-Alba C., Kim C.F., Gomperts B., Arumugaswami V, & Stripp B.R. (2021). SARS-CoV-2 infection of primary human lung epithelium for COVID-19 modeling and drug discovery. Cell Reports, 35(5), 109055.

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Berberine-Sonodynamic Therapy for Macrophages

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BBR was obtained from Chengdu Must Bio-Technology Co. (Chengdu, China) and was stored in ddH₂O as a 1 mg/ml stock solution at 4 °C in the dark. When THP-1 macrophages reached an exponential phase, they were collected, differentiated, and randomly divided into four groups: (1) control, (2) ultrasound alone, (3) berberine alone, and (4) BBR-SDT. For the berberine and BBR-SDT groups, the cells were incubated with the indicated doses of BBR for 4 h in RPMI 1640 medium containing FBS. The control and ultrasound alone groups received an equivalent volume of medium instead of BBR. The cells in the ultrasound and BBR-SDT groups were exposed to ultrasound at a frequency of 1.0 MHz for the indicated intensities. After the treatments, the cells were carefully washed once in phosphate-buffered saline (PBS), were cultured in fresh medium for a few hours, and then were subjected to different analyses.
Depending on the experiments performed, 3MA (Sigma-Aldrich), LY294002 (Selleck Chemicals, Houston, TX, USA), rapamycin (Rapa; Selleck Chemicals), or hydroxychloroquine (Selleck Chemicals) were added to the culture medium in combination with BBR for 4 h. When appropriate, NAC was added to the culture medium 30 min prior to the addition of BBR-SDT.

Kou J.Y., Li Y., Zhong Z.Y., Jiang Y.Q., Li X.S., Han X.B., Liu Z.N., Tian Y, & Yang L.M. (2017). Berberine-sonodynamic therapy induces autophagy and lipid unloading in macrophage. Cell Death & Disease, 8(1), e2558-.

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Cellular Metabolism Profiling Protocol

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The antibodies of NOX2(ab129068), COX-2, NOX-1, GAPDH and β-actin were purchased from abcam (Cambridge, MA, USA); the antibodies of AKT(pan), phospho-AKT(Ser473), LC3B, phospho-AMPK and AMPK were purchased from Cell Signaling Technologies (Beverly, MA, USA); the antibodies of MCT4(D-1)(sc-376140) , GLUT1, GLUT3, goat anti-rabbit and goat anti-mouse were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). 2-DG(2-deoxy-D-glucose) (D109194), oligomycin (O111756), nevirapine (N129779), glucose, ATP, ADP and AMP were purchased from Aladdin chemicals (Shanghai, China). PMS(P9625) was purchased from Sigma-Aldrich (Shanghai, China); MTS Reagent (G1112) was purchased from Promega Corporation (WI, USA). Seahorse XF Cell Test Kit was purchased from Seahorse Agilent technologies (Beijing, China). RPMI-1640(8116322) and glucose-free RPMI 1640(1779211) were purchased from Gibco Thermo Fisher Scientific (Guangzhou, China). Hydroxychloroquine, MK2206 and Compound C were purchased from Selleck Chemicals (Houston, TX, USA). GSH/GSSG and ROS test kits were purchased from Beyotime Biotechnology (Shanghai, China). Acetonitrile and methanol (HPLC grade) were purchased from Oceanpak (Goteborg, Sweden).

Ye M., Wang S., Wan T., Jiang R., Qiu Y., Pei L., Pang N., Huang Y., Huang Y., Zhang Z, & Yang L. (2017). Combined Inhibitions of Glycolysis and AKT/autophagy Can Overcome Resistance to EGFR-targeted Therapy of Lung Cancer. Journal of Cancer, 8(18), 3774-3784.

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BA Regulation of Signaling Pathways

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BA (purity >97%) was purchased from Aladdin (Shanghai, China) and dissolved in DMSO. A stock solution of 10 mM BA was prepared and stored at −20°C. SP600125 (JNK inhibitor), SB203580 (p38 inhibitor), U0126 (ERK inhibitor), MK2206 (AKT inhibitor), hydroxychloroquine (HCQ, autophagy inhibitor), trametinib, and MEK162 (ERK inhibitor) were obtained from Selleck Chemicals (Houston, United States). Primary antibodies follow: β-actin (CST, 4970S), GAPDH (Abcam, ab181602), LC3 (CST, 12741S), ATG5 (CST, 12994S), p38 (Santa Cruz, sc-271120), p-p38 (Santa Cruz, sc-166182), ERK1/2 (Proteintech, 16443-1-AP), p-ERK1/2 (ST, 4370T), JNK (Santa Cruz, sc-7345), p-JNK (CST, 4668T), PARP (CST, 9542S), cleaved PARP (CST, 5625S), AKT (CST, 4691S), p-AKT (CST, 4060P), CDK2 (Abcam, ab205718), CDK4 (Abcam, ab108357), and cyclin D1 (Abcam, ab134175). Horseradish peroxidase (HRP)-conjugated secondary antibodies were supplied by Cell Signaling Technology.

Sun C.Y., Cao D., Ren Q.N., Zhang S.S., Zhou N.N., Mai S.J., Feng B, & Wang H.Y. (2021). Combination Treatment With Inhibitors of ERK and Autophagy Enhances Antitumor Activity of Betulinic Acid in Non–small-Cell Lung Cancer In Vivo and In Vitro. Frontiers in Pharmacology, 12, 684243.

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KRAS siRNA and shRNA Knockdown Assay

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siRNA oligonucleotides targeting KRAS were: KRAS 1, CUAUGGUCCUAGUAGGAAAtt (Thermo Fisher, 4390824, s7939), and KRAS 2, GCCUUGACGAUACAGCUAAtt (Thermo Fisher, 4390824, s7940). The target sequence for the validated shRNA construct used to target KRAS was CAGTTGAGACCTTCTAATTGG. To determine the levels of activated proteins, immunoblot analyses were done using phosphospecific antibodies to AMPKα (T172; 40H9), Beclin-1 (S93; D9A5G), DRP1 (S616) and RSK (T395/S363). Antibodies recognized total LC3B, KRAS (234–4.2), AMPKα (Δ63Γ4) Beclin-1 (2A4), PINK1 (D8G3), VDAC (D73D12), DRP1 (D8H5), RSK (32D7), ATG5, ATG7 (EP1759Y), β-actin (AC-15), vinculin (hVIN-1) or GAPDH (GAPDH-71.1) to control for total protein expression. All antibodies were obtained from Cell Signaling Technologies except ATG7 (Abcam), KRAS (Calbiochem), β-actin (Sigma) and vinculin (Sigma). Immunochemical labeling of mitochondria was performed using TOMM20 (Santa Cruz). Chemical reagents used were bafilomycin A1, oligomycin A, rotenone, antimycin, FCCP, CCCP, doxycycline, MTT and chloroquine diphosphate (Sigma); calcein AM and AlamarBlue (Invitrogen); hydroxychloroquine (SelleckChem); binimetinib (SelleckChem); ARS-1620 (Medchem Express); Spautin-1, SBI-0206965 and MRT68921 (Xcessbio); SCH772984 (provided by Merck) and BVD-523 (provided by Biomed Valley Discoveries).

Bryant K.L., Stalnecker C.A., Zeitouni D., Klomp J.E., Peng S., Tikunov A.P., Gunda V., Pierobon M., Waters A.M., George S.D., Tomar G., Papke B., Hobbs G.A., Yan L., Hayes T.K., Diehl J.N., Goode G.D., Chaika N.V., Wang Y., Zhang G.F., Witkiewicz A.K., Knudsen E.S., Petricoin EF I.I.I., Singh P.K., Macdonald J.M., Tran N.L., Lyssiotis C.A., Ying H., Kimmelman A.C., Cox A.D, & Der C.J. (2019). Combination of ERK and autophagy inhibition as a treatment approach for pancreatic cancer. Nature medicine, 25(4), 628-640.

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