Integrated and Functional Genomics Analysis Validates the Relevance of the Nuclear Variant ErbB380kDa in Prostate Cancer Progression
Fig 5
(A) Two siRNAs were used to specifically inhibit the NM_001982.3 transcript (siErbB3A) or both the NM_001982.3 and AK300909.1 transcripts (siErbB3B) and validated by western blotting using the ErbB3 C-ter antibody. (B) LNCaP and PC3 cells were transiently transfected with siErbB3A or siErbB3B before mRNAs were reverse transcribed and amplified. RT-qPCR was performed on a selection of ErbB3-target genes. Expression fold change was normalised to control siRNA transfected samples. Transfections were done in triplicate, and the RT-qPCR results reported are from at least three independent experiments. (C) Expression fold change consecutive to the inhibition of the ErbB380kDa isoform in each cell line and for each target gene is calculated by the ratio (relative gene expression upon siErbB3B treatment)/(relative gene expression upon siErbB3A treatment). ErbB380kDa-dependent transcriptional activation of GATA2, RUNX2, MAP3K14, ErbB2 and CCND1 and inactivation of PPIG, MXI1 were similar in LNCaP and PC3 whereas ARID1A, TBCC, FYN, SIN3A, TOR3A, ACE, RAD52, CXCR3A appeared to be differentially regulated in the two cell lines. *P<0.05, **P<0.01, ns = non-significant, Student’s t test.