Fig. 6

Knockdown of ZNF507 leads to altered tumor growth in vivo and diminish NEPC-like phenotypes by TGF-β signaling inhibition. A Growth curve of the tumor formation after the scramble, shZNF507, Mock, or ZNF507OE DU145 cells were xenografted into Balb/C nude mice (n = 6 per each set group). For each group, three tumor samples were used for the Western blot analysis, while other three samples were used for the IF. B Tumors from scramble and shZNF507 group were extracted on the last day (day 33) and Mock and ZNF507 OE group were extracted on day 18 of the xenograft experiments conducted through injection. C Graph showing the average weight of the scramble and shZNF507 xenografted tumors (n = 6 per each group). D Histological analysis of tumors from the scramble and shZNF507 xenograft. Representative data of H&E staining and immune staining against ZNF507 (with PI), TGFBR1, MAP3K8, and FURIN in tumors, performed by immunofluorescence (Scale bar = 100 μm). E Representative protein expression data of ZNF507, TGFBR1, MAP3K8, and FURIN in the xenografted tissues as determined by western blot analysis. β-actin served as a loading control. Right panel shows the average of the relative protein levels normalized to the β-actin. At least three samples per group were used for each experiment and the three independent experiments were performed and analyzed. F Representative protein expression data of canonical TGF-β signal proteins, Smad2, Smad3, and their phosphorylated forms, assessed by western blot. β-actin was used as a normalization control. At least three samples per group were used for each experiment and the three independent experiments were performed and analyzed. G Representative protein expression levels of non-canonical TGF-β signal proteins, RAS, MEK, and p-MEK, ERK, and p-ERK, examined by western blot. β-actin was used as a normalization control. At least three samples per group were used for each experiment and the three independent experiments were performed and analyzed. H Representative data of H&E staining of the liver and lung from in vivo metastasis assay. I Representative images of the immune staining against ZNF507, TGFBR1, MAP3K8, and FURIN in liver and lung from in vivo metastasis assay, performed by immunofluorescence. J Schematics presenting the biological role of ZNF507 in TGF-β signaling, as a transcriptional activator, and related pathway in the progression of PC to its aggressive mCRPC and NEPC state. Data are presented as the Means ± SD from three independent experiments. #p < 0.05, ##p < 0.01, Mock versus the ZNF507 OE xenograft tissue and *p < 0.05, **p < 0.01, ***p < 0.001 shZNF507 versus the scramble xenograft tissue