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. 2021 Jan 8;296:100254. doi: 10.1074/jbc.RA120.015769

Figure 6.

Figure 6

Expression of exogenous TRPC6 reverses the antiproliferative effect of melatonin in MDA-MB-231 cells. MDA-MB-231 cells were treated for 48, 72, and 168 h with melatonin (10–1000 nM) or the vehicle (control). Forty-eight hours before the end of the treatment period, cells were transfected with TRPC6-YFP (B, E, and H) or TRPC6dn-YFP (C, F, and I) expression plasmid or empty vector (A, D and G). Cell proliferation was assessed at time = 0, 24, 48, and 72 h using the BrdU cell proliferation assay kit, as described in Experimental procedures. Scatter plots represent cell proliferation 0, 24, 48, and 72 h after the onset of the experiment, presented as BrdU uptake rate (n = 6). Analysis of statistical significance was performed using two-way ANOVA (for A, D, G: 0 h, F values were 0.14, 0.14, and 1.35 and p values were 0.93, 0.86, and 0.24 for concentration, time, and the interaction respectively; for A, D, G: 24 h, F values were 593.8, 1570, and 199.6 and p values were <0.0001 in all cases; for A, D, G: 48 h, F values were 512.2, 1309, and 201.4 and p values were <0.0001 in all cases; for A, D, G: 72 h, F values were 1332, 5690, and 619.2 and p values were <0.0001 in all cases; for B, E, H: 0 h, F values were 0.03, 0.0.05, and 0.03 and p values were 0.99, 0.94, and 0.99; for B, E, H: 24 h, F values were 0.09, 0.09, and 0.05 and p values were 0.90, 0.98, and 0.99; for B, E, H: 48 h, F values were 0.09, 0.10, and 0.11 and p values were 0.89, 0.98, and 0.99; for B, E, H: 72 h, F values were 0.15, 0.05, and 0.07 and p values were 0.96, 0.94, and 0.99; for C, F, I: 0 h, F values were 0.05, 0.22, and 0.02 and p values were 0.99, 0.79, and 0.99; for C, F, I: 24 h, F values were 78.10, 2894, and 66.80 and p values were <0.0001 in all cases; for C, F, I: 48 h, F values were 704,6, 2442, and 169 and p values were <0.0001 in all cases; for C, F, I: 72 h, F values were 893.70, 3707, and 240.1 and p values were <0.0001 in all cases) with post-hoc Tukey's test for comparison between the data corresponding to the same time of evaluation of proliferation (0, 24, 48, or 72 h) (∗p < 0.0001 compared with the corresponding control [cells not treated with melatonin]).