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. 2020 Apr 16;9(4):989. doi: 10.3390/cells9040989

Figure 4.

Figure 4

Tumor acidosis-mediated decrease of BMAL1 expression promotes metastatic potency in breast cancer cells. (a) The green fluorescent protein (GFP) or GFP-BMAL1 stably overexpressed MDA-MB-231 cell lines were incubated in HCl-mediated acidic condition for 24 h as indicated, and cell lysates were determined by immunoblotting. (b,c) GFP or GFP-BMAL1 stably overexpressed MDA-MB-231 cell lines were subjected to trans-well migration assay in HCl-mediated acidic condition for 24 h as indicated. Representative images of migrated cells are shown. Scale bars: 250 μm (b). The average number of migrated GFP or GFP-BMAL1 stably overexpressed MDA-MB-231 cell lines was counted in three random microscopic fields (c). (d) MDA-MB-231 was transfected with si-NTC or si-BMAL1. Cell lysates were determined by immunoblotting. (e,f) MDA-MB-231 was transfected with si-NTC or si-BMAL1, and was subjected to trans-well migration assay for 24 h. Representative images of migrated cells are shown. Scale bars: 250 μm (e). The average number of migrated MDA-MB-231 was counted in three random microscopic fields (f). Data represent the mean ± SD, n = 3. ** p < 0.01 and *** p < 0.001 vs. the control group or between two groups by a Student’s t-test.