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. 2017 Jul 14;7:5506. doi: 10.1038/s41598-017-05421-x

Figure 1.

Figure 1

Ssu72 regulates STAT3 activation. (A) NIH-3T3 cells were transfected with either the mock or Ssu72 overexpression vector and stimulated with IL-6 (20 ng/ml) for 1 h. Cells were used to detect the p-STAT3 Tyr705 and p-STAT3 Ser727 levels. (B) NIH-3T3 cells were stained with DAPI (blue) and antibodies against tubulin (green) and p-STAT3 Tyr705 (red). Confocal scanning microscopy was performed to detect the p-STAT3 Tyr705 and p-STAT3 Ser727 levels in the transfected cells. (C) NIH-3T3 cells were transfected with the expression vector and stimulated with IL-6 (20 ng/ml) for 1 h. (D) Cells were transfected with STAT3 overexpression vectors and either the mock or Ssu72 overexpression vector. The expression levels of the Il17a mRNA were measured using real-time PCR. (E) NIH-3T3 cells were transfected with the Il17a promoter construct and either mock or Ssu72 expression vectors. Luciferase activity was then detected. (F) Lysates from the transfected NIH-3T3 cells were immunoprecipitated with the anti-FLAG antibody and immunoblotted with anti-p-STAT3 Tyr705, anti-p-STAT3, and anti-Ssu72 antibodies. The data represent the mean ± SD from three independent experiments. Statistical analyses were conducted using the nonparametric Mann-Whitney U-test (*P < 0.05, ***P < 0.01).