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. 2017 May 18;7:2090. doi: 10.1038/s41598-017-02242-w

Figure 5.

Figure 5

Sp3 but not Sp1 regulated the mSTING proximal promoter activity. (A,B) Overexpression of Sp1 or knockdown of Sp1 had no effect on the activity of the mSTING promoter. While knockdown of Sp3 by siRNA reduced the mSTING promoter activity to 35%, and overexpression of Sp3 increased the promoter activity by more than 2 folds. NIH3T3 cells were co-transfected with Sp1/Sp3 siRNA (50 nM) or Sp1/Sp3 expressing plasmid (200 ng) and pSTING-254(100 ng) and inner control pRL-TK (3 ng). After 24 h, luciferase activity was measured. (C,E,G) Reduction of endogenous Sp3 expression by siRNA decreased expression of the mSTING gene at mRNA and protein level. qRT-PCR and western blot were performed after siRNA/NC(50 nM) were transiently cotransfected into NIH3T3 cells. siRNA negative control was set as 1. Each bar represented the mean ± SD of three independent experiments. (n = 3, *P < 0.05 vs.NC). (D,F,H) STING protein expression increased by two times under overexpression of Sp3. Sp3 plasmid, pcDNA 3.1(+) (4000 ng) were transiently cotransfected into NIH3T3 cells, and then qRT-PCR and western blot were performed. Each bar represented the mean ± SD of three independent experiments. (n = 3, *P < 0.05 vs. Control).