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. 2021 Jan 12;13(2):259. doi: 10.3390/cancers13020259

Figure 4.

Figure 4

SPCA2 is required for HDACi-induced MET changes in TNBC cell lines. (A) Schematic showing that HDAC inhibitors regulate SPCA2 expression, which in turn, promotes MET. (B) SPCA2 mRNA, determined by qPCR, shows efficacy of knockdown (KD) in MDA-MB-231 cells relative to scramble shRNA control, and (C) following treatment with vehicle (control) or vorinostat (2.5 µM) for 24 h, as indicated. (D–F) Expression of mesenchymal gene markers N-cadherin, vimentin and ZEB1 was determined by qPCR following treatment with vehicle (control) or vorinostat (2.5 µM). SPCA2KD effectively reversed decrease in mesenchymal gene expression in the presence of vorinostat. n = 3. (G) Representative microscope images of MDA-MB-231 cells and MDA-MB-231 SPCA2 KD treated with vehicle or vorinostat (2.5 µM) for 24 h, in Boyden chamber (10× magnification). (H) Migration was quantified by ImageJ software from 3 images for each condition. (I) SPCA2 KD in MDA-MB-231 significantly reduced cytotoxic effect of vorinostat (n = 6, 2.5 μM for 80 h, MTT assay). Student t-test, *** p < 0.001, ** p < 0.01, * p < 0.05.