Figure 6. Knockdown of IL-13RA2 in NFs induced keloid-characteristic behaviors via JAK/STAT6.

(A) Growth curves of NFb cells upon IL-13RA2 knockdown by CCK8 assay. NC, non-treatment control. (B) Annexin V/7-AAD double staining assay of NFb cells upon IL-13RA2 knockdown with or without AS1517499 treatment. NFb cells were treated with or without 1 μM AS1517499 for 16 hours before Annexin V/7-AAD double staining. (C and D) Migration and invasion of NFb upon IL-13RA2 knockdown with or without AS1517499 treatment. Cells (3 × 10⁴) were seeded in upper Transwell chambers and treated with or without 1 μM AS1517499 for 16 hours for the migration assay, while 6 × 10⁴ cells were seeded and treated with or without 1 μM AS1517499 for 22 hours for the invasion assay. Quantification of migrated and invaded cells is shown on the right. Scale bars: 100 μm. (E) Representative images of α-SMA immunofluorescent staining in NFb cells upon IL-13RA2 knockdown. Scale bars: 100 μm. (F) Western blot results of NFb cells upon IL-13RA2 knockdown. NFb cells were treated with or without 1 μM AS1517499 for 16 hours and then analyzed by Western blotting. Relative gray scales of p-STAT6/STAT6 and indicated protein/GAPDH are shown below the blots. Error bars represent SD. Experiments were performed 3 times and in triplicate each time. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 by 2-tailed Student’s t test. NS, not significant (P > 0.05).