Bafilolycin a1

Manufactured by InvivoGen

Bafilomycin A1 is a macrolide antibiotic that acts as a selective and potent inhibitor of vacuolar-type H+-ATPase (V-ATPase). V-ATPase is responsible for acidifying various intracellular compartments, such as endosomes, lysosomes, and secretory vesicles. Bafilomycin A1 inhibits this proton pump, leading to the disruption of pH homeostasis within these organelles.

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Lab products found in correlation

2 protocols using bafilolycin a1

Recombinant Protein-Mediated Cell Signaling

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Recombinant human TGF-β, recombinant FGF-2 and recombinant human TNF-α were purchased from Biolegend (San Diego, CA). Rapamycin was from ENZO life science (Loerrach, Germany). Bafilolycin A1 was from Invivogen. FITC-PHA was from Vector (FL-1111). FITC-PNA was from Sigma (L7381). DAPI was from Sigma (D9542). Alexa Flour 488, 594 or 647 conjugated anti-mouse or anti-rabbit IgG were from Jackson ImmunoResearch Laboratories (West Grove, PA). The primary antibodies against JLP (ab12331, 1:1000), Fsp-1 (ab197896, 1:200), α-SMA (ab124964, 1:1000), Fibronectin (ab45688, 1:500), Collagen-I (ab34710, 1:1000), Ki67 (ab16667, 1:250), TGF-β (ab92486, 1:500), phospho-Smad2 (ab188334, 1:1000), phospho-Smad3 (ab52903, 1:1000) and p62 (ab56416, 1:200) were all from Abcam (Cambridge, MA). Anti-LC3 antibody (ab51520, 1:100, Abcam) was used for immuno-staining and Anti-LC3 antibody (L7543, 1:1000, Sigma) was used for western blotting, respectively. Anti-Nephrin and anti-F4/80 (123120, 1:100) antibodies were from Progen and Biolegend, respectively. Anti-Caspase-3 (#9664, 1:1000), anti-Beclin 1 (#3738,1:1000) was from Cell signaling Technology (Danvers, MA). Anti-GAPDH (sc-365062, 1:2500) was purchased from Santa Cruz (Santa Cruz, CA).

Yan Q., Zhu K., Zhang L., Fu Q., Chen Z., Liu S., Fu D., Nakazato R., Yoshioka K., Diao B., Ding G., Li X, & Wang H. (2020). A negative feedback loop between JNK-associated leucine zipper protein and TGF-β1 regulates kidney fibrosis. Communications Biology, 3, 288.

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Evaluating TGF-β, FGF-2, and TNF-α effects on HK-2 cells

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Human TEC cell lines HK-2 was purchased from China Centre for Type Culture Collection and was maintained as previously described. Recombinant human TGF-β, recombinant FGF-2 and recombinant human TNF-α were purchased from Biolegend (San Diego, CA). HK-2 cells were treated with TGF-β or recombinant FGF-2 or recombinant human TNF-α for 24 h, respectively, and then were harvested and analyzed by qPCR and western blotting. Jlp siRNA and control siRNA was purchased from Qiagen (Germany). The siRNA transfection was carried out using HiPerFect transfection (Qiagen) according to customer’s protocol. The efficiency of transfection was assessed by the protein expression. The sequence used for knockdown of Jlp in this study was: 5′-CAGACCCGAGTGGAATCTTTA-3′. Bafilolycin A1 was from Invivogen. HK-2 cells treated with or without Bafilolycin A1, as well as the cells transfected with Jlp siRNA for 24 h,

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