Fig. 1: PBRM1/Pbrm1 deficiency reduced IFNγ-STAT1 activity in Renca cells and 786-O cells. | Nature Communications

Fig. 1: PBRM1/Pbrm1 deficiency reduced IFNγ-STAT1 activity in Renca cells and 786-O cells.

From: PBRM1 loss defines a nonimmunogenic tumor phenotype associated with checkpoint inhibitor resistance in renal carcinoma

Fig. 1: PBRM1/Pbrm1 deficiency reduced IFNγ-STAT1 activity in Renca cells and 786-O cells.The alternative text for this image may have been generated using AI.

a Pbrm1 knockout validation in Renca cells at protein levels by western blot, and b at mRNA levels by real-time PCR. Renca cell were treated with or without 1 ng/ml IFNγ for 8 h. c IFNγ-induced JAK-STAT1 expression and phosphorylation in Renca cells. Control KO or Pbrm1 KO (clone #18) Renca cells were treated with 1 ng/ml IFNγ for 2 or 8 h. Cell lysates were analyzed by immunoblot using antibodies against PBRM1, STAT1, P-STA1 Y701, P-STAT1 S727, JAK2, P-JAK2 Y1007/1008, JAK1, P-JAK1 Y1034/1035, IRF1. β-actin was used an internal control. d IFNγ-induced gene expression in Renca cells. Control KO or Pbrm1 KO (clone #18) Renca cells were treated with 1 ng ml IFNγ for 8 h. mRNA expression of Stat1, Cxcl9, Irf1, and Icam1 were detected by real-time PCR. Gapdh was used as internal control. e IFNγ-induced CXCL9 secretion. Renca cells were cultured in serum-free medium and treated with 1 ng/ml IFNγ for 4 or 10 h. The concentration of CXCL9 was analyzed using Quantikine® ELISA kit. f IFNγ-induced JAK-STAT1 expression and phosphorylation in 786-O cells. Control knockdown (Con KD) or PBRM1 knockdown (PBRM1 KD) 786-O cells were treated with or without 10 ng/ml IFNγ for 2 h. Cell lysates were analyzed by immunoblot using antibodies against PBRM1, STAT1, P-STA1 Y701, JAK2, P-JAK2 Y1007/1008, and IRF1. β-actin was used an internal control. g IFNγ-induced gene expression in 786-O cells. 786-O cells were cultured in DMEM with 10% FBS, and treated with 10 ng/ml IFNγ for 8 h. mRNA expression of STAT1, CXCL9, and IRF1 were detected by real-time PCR. GAPDH was used as internal control. Unpaired t-test was performed with GraphPad Prism 7.03. *P < 0.05 and **P < 0.001, compared with control knockout or knockdown cells. All data are representative of three independent experiments. Data in the bar graphs represent mean ± S.D., n = 3. Source data are provided as a Source Data file.

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