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. 2019 Sep 2;20(17):4293. doi: 10.3390/ijms20174293

Figure 5.

Figure 5

Combination of hnRNPA1 knockdown and JQ1 decreases Survivin. (A) CD18 pancreatic cancer cells were transfected with control siRNA (siCtrl) or hnRNPA1-targeting siRNA (sihnRNPA1, siA1) for 48 h. The cells were then treated with DMSO or JQ1 (1 µmol/L) for an additional 24 h. Cell lysates were collected and analyzed for apoptosis-related proteins using Proteome Profiler Human Apoptosis Array ARY009, and the pixel density of Survivin from the array data, highlighted with white boxes, was quantified by ImageJ. hnRNPA1 knockdown was confirmed by western blot analysis. Error bars represent SD from two technical replicates. The results are representative of two independent experiments. (B) Cancer cells transfected with siCtrl or sihnRNPA1 (siA1) for 48 h were subsequently treated with DMSO or JQ1 (1 µmol/L) for an additional 24 h. Cell lysates were analyzed for expression of hnRNPA1 and Survivin by western blot analysis, using HSP90 as loading control. (C) Average pixel density from two independent experiments with CD18 cells was calculated for each protein in the array and normalized to the corresponding control treatment group. Heat maps reflect % change in the expression of select proteins in cells transfected with siCtrl or sihnRNPA1 (siA1) and co-treated with JQ1 and in cells treated with JQ1 alone or a combination of JQ1 and Quercetin (Quer). *, proteins common in both heat maps. DMSO: dimethyl sulfoxide; JQ1: BET inhibitor; HSP90: heat shock protein 90 (loading control); hnRNPA1: heterogeneous nuclear ribonucleoprotein A1; cIAP-2: cellular inhibitor of apoptosis protein 2; HSP70: heat shock protein 70; SD: standard deviation; K1: human papillary thyroid cancer cell line; 8505c: human anaplastic thyroid cancer cell line; CD18: human pancreatic cancer cell line.