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. 2016 Sep 8;17(9):1504. doi: 10.3390/ijms17091504

Figure 3.

Figure 3

LiCl regulates the expression of Six2 at mRNA and protein level. (A) pGL3-Six2 promoter-Luciferase construction was simulate by diagram. The Six2 promoter ranging from −2322 to −323 (Six genome sequence) was obtained from NCBI; (B) HEK293T cells were co-transfected with pRL-SV40 (renilla control) and pGL3-Six2 promoter-LuC for 36 h. Luciferase activity was normalized to Renilla control. p-values were calculated by Student t-test. Values represents mean values ± SEM of triplicate experiments, *** p < 0.001 relative to control; (C) HEK293T cells were co-transfected with pRL-SV40 (renilla control) and pGL3-Six2-LuC for 36 h then were treated with LiCl of increasing dosages for 12 h. Luciferase activity was measured using dual luciferase reporter assay, normalized to Renilla control. Values were presented as mean ± SEM (n = 3), *** p < 0.001 relative to control; (D) mK3 cells were treated with LiCl of increasing dosages for 12 h. The Six2 expression at protein level was tested by Western-blot. Values were presented as mean ± SEM (n = 3), ** p < 0.01 relative to control.