Figure 4
From: Distinct outcomes of CRL–Nedd8 pathway inhibition reveal cancer cell plasticity
Distinct roles of CRL components in cell regulation. (a) Effect of gene knockdown on VCaP cell proliferation and survival was measured after 5 days of siRNA treatment and compared with control siRNA (siCTL) and 50 and 500 nM MLN (MLNlow and MLNhigh). Cell number and ATP level were normalized to the values of siCTL-treated cells. The major ‘hits’ are shown (for all genes, see Supplementary Figure S9). (b) Survival of prostate cancer cells after RBX1 knockdown with two individual siRNAs. Statistical robust Z-score was used (RZ, see Supplementary Information). Knockdown efficacy was confirmed by western blotting with Rbx1-specific antibody. (c) Survival of prostate cancer cells upon CAND1 knockdown with two individual siRNAs. Analysis as in b. (d) Effect of CAND1 knockdown on MLN-induced apoptosis. VCaP cells were transfected with 5 nM of CAND1 siRNA or with the same amount of siCTL and treated with MLN (see also Supplementary Figure S10). (e) CAND1 transcripts are upregulated in prostate cancer. Expression of CAND1 in benign prostatic hyperplasia (BHP, blue), prostatic intraepithelial neoplasia (PIN, blue), and prostate cancer (PCa, blue) in comparison with normal prostatic tissue (gray) (Oncomine database,27 www.oncomine.org). (f) Immunohistochemical staining for Cand1 in prostate tumors and normal tissues (Human Protein Atlas,28 www.proteinatlas.org)
