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. 2016 Apr 9;7(20):29063–29080. doi: 10.18632/oncotarget.8666

Figure 2. Silencing of sM8 isoforms reduces cell growth of prostate cancer cell lines and triggers stress in endoplasmic reticulum.

Figure 2

(A) Cell growth of LNCaP, LNCaP C4-2b and PC-3 cells was measured with CellTiter 96® AQueous Non-Radioactive Cell Proliferation Assay (Promega), after 2 or 3 days of culture. Average values at day 2 and 3 were normalized to values at D0. Experiments were performed three times independently. (B) Kinetic of LNCaP C4-2b cell growth following an identical procedure than the one described in A. Cells were transfected with either control siRNA (siLuc) or anti-TRPM8 siRNA (siM8-6a, siM8-7, siM8-20), labels refer to the target TRPM8 exon. A single point mutant siM8-6a showing a limited efficiency, siM8-6a(M1), demonstrates the specificity of the siM8-6a sequence on sM8 KD-mediated inhibition of cell growth. Experiments were performed four times independently. (C) Gene expression was estimated with real time PCR after a 3-day siRNA transfection of LNCaP C4-2b cells. Graph plot displays human genes involved in endoplasmic reticulum stress: ATF4, ATF6, HSPA5, DDIT3 (Chop), EIF2AK3 (PERK), HSP60 and XBP1. Experiments were performed three times independently. Values are expressed as Mean ± SD.