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. 2017 Apr 11;8(24):39417–39429. doi: 10.18632/oncotarget.17034

Figure 6. Loss of STAT3 inhibition by edited BLCAP after YXXQ motif mutation.

Figure 6

(A) The coding sequence of wild type BLCAP and two mutants. A>G substitutions (A-to-I RNA editing) in DNA sequence were oversized. Letters under the line represented the amino acid translated by DNA sequence underlined. Three types of BLCAP recombinant plasmids were transfected into HeLa cells and harvested after 24 hours, then Western blot was conducted with anti-FLAG antibody. (B) The wild type BLCAP and two mutants were transfected into HeLa cells respectively, followed by immunoprecipitation with an anti-FLAG antibody and immunoblot analysis with the indicated antibodies. (C) PCAGGS and three plasmids of BLCAP were transfected into HeLa (left) and C33A (right) cells for 24 hours. HeLa cells were harvested after IL-6 (10ng/ml) stimulated for 10 minutes and C33A cells were lysed after IL-6 (100ng/ml) stimulated for 4 minutes. The whole-cell lysates were processed for Western blot with the indicated antibodies. β-actin was used as a loading control. (D) PCAGGS and three types of BLCAP plasmids were transfected into HeLa cells for 24 hours to perform realtime-PCR assay. Data were shown by analysis of unpaired Student's t test, ***P<0.001.