Fig. 3
From: EGFR feedback-inhibition by Ran-binding protein 6 is disrupted in cancer
RanBP6 promotes nuclear translocation of STAT3. a RanBP6 knockout (KO) increases cytoplasmic STAT3 and lowers nuclear STAT3. Left panel, immunoblots of cytoplasmic (Cyt.) and nuclear (Nuc.) cell fractions; right panel, densitometry of STAT3 immunoblots. b RanBP6 KD impairs IL-6-induced nuclear STAT3 translocation. Left panel, confocal immunofluorescence. RFP is used as a reporter for shRNA expression. Right panel, ratios of nuclear/cytoplasmic STAT3 staining (field of views: vehicle, n = 18; ruxolitinib, n = 13; RANBP6-shRNA, n = 21). The janus kinase (JAK) inhibitor ruxolitinib was included as a positive control. Scale bar = 10 μm. c RanBP6 KD decreases transcription of STAT3 reporter gene. SIE sis-inducible elements. d Gene expression profiling showing the effect of RanBP6 KD on endogenous STAT3 target genes. Heatmap represents the enrichment scores from single-sample gene set enrichment analysis (ssGSEA) of three biological replicates. Student’s t test p-values (Dox− vs. Dox+) for each gene sets are indicated. e Quantitative PCR analysis of the expression of some RanBP6-regulated genes selected from ssGSEA (top panel) confirmed to be regulated by STAT3 (bottom panel). Data are represented as mean ± SD (n ≥ 3). Student’s t test: ***p < 0.001; **p < 0.01; ns not significant
