Figure 5
FAM83A promotes activation of pro-survival MEK/ERK signalling. RNA sequencing analysis of RNA extracted from HPAC-shGFP, HPAC-shA2 and HPAC-shA6 cells shown as heatmaps (a) indicated reduced expression of poor prognosis markers of pancreatic cancer and increased expression of pro-apoptotic genes. Data from each triplicate sample are shown. (b) qRT-PCR for pro-apoptotic Bid and immunoblotting for cleaved caspase-3 at 96 h (c). The control in (c) is lysate from HPAC cells treated with etoposide to induce cleaved caspase-3. (d) Pro-survival gene signature shown as heatmap was extracted from the RNA sequencing experiment described in (a). (e) qRT-PCR analysis for ERK- transcriptional targets EGR1, JUNB and FOSB. Significance values shown are relative to the shGFP control group. (f) Activation of MEK and PI3K pathways were assessed in HPAC-shGFP and HPAC-shA2 cells by measuring phosphorylation of ERK, mTOR and AKT. GAPDH was used as control. Data in C and F are representative of three independent experiments. Student’s t-test (two-tailed, unequal variance) was used to assess statistical significance with *, ** and *** representing p < 0.05, p < 0.01 and p < 0.001 respectively.
