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Fig. 6 | Cell Communication and Signaling

Fig. 6

From: Exosomal AP000439.2 from clear cell renal cell carcinoma induces M2 macrophage polarization to promote tumor progression through activation of STAT3

Fig. 6

AP000439.2 interacts with STAT3 and phosphorylates STAT3 in macrophage. A AP000439.2 was located dominantly in the nucleus proved by cell fractions isolation followed with qRT-PCR. GAPDH was set as a cytoplasm control, and U6 was set as a nucleus control. B STAT3 was detected in the antisense and AP000439.2 RNA pull-down sample by western blot. RNA pull-down was performed in macrophages. Antisense RNA probe was used as negative control for AP000439.2 probe. C RNA immunoprecipitation with an anti-STAT3 antibody was used to assess whether STAT3 binding to AP000439.2 in macrophages. IgG antibody was used as the control. D Deletion mapping of the AP000439.2-binding domain in STAT3. Left, predictive secondary structure of AP000439.2; Up, diagrams of full-length AP000439.2 and the deletion fragments; Bottom, western blot analysis for STAT3 antibody in samples pulled down by the different AP000439.2 constructs. E Deletion mapping of the STAT3-binding domain in AP000439.2. Up, diagrams of full-length STAT3 and the deletion fragments; Bottom, western blot analysis for AP000439.2 probe in samples pulled down by the different STAT3 constructs. F The phosphorylation level of STAT3 in macrophages decreased upon AP000439.2 knocking down compared with the negative control. G The protein expression level of TGF-β and IL-10 in macrophage was detected by western blot after overexpression of AP000439.2 and knockdown of STAT3. **p < 0.01, ***p < 0.001, Two-tailed Student’s t-test for two groups, one-way ANOVA following Tukey’s test for three groups

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