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. 2020 Jul 9;10:11388. doi: 10.1038/s41598-020-68219-4

Figure 6.

Figure 6

IKK-β and IKK-α are not involved in TNF-α induced phosphorylation of SRC-3 at S857. (A) IKK-β does not phosphorylate SRC-3 at S857. A549 cells pretreated with either DMSO or 1, 5, 10, 20 μM BI-605906 for 30 min were stimulated with 10 ng/ml TNF-α for 15 min. Then the cells were lysed and Western-blotting was carried out to examine the phosphorylation of SRC-3 at S857, HSP27 at S82 and expression of total SRC-3, HSP27, IκBα using anti-P-S857-SRC-3, anti-P-HSP27, anti-SRC-3, anti-HSP27 and anti-IκBα antibodies respectively. The full-length blots are presented in supplementary figure S33. (B) IKK-α does not phosphorylate SRC-3 at S857. A549 cells were transfected with either 10 nM scrambled siRNA, siRNA against IKK-α or left untransfected. After 48 h, the cells were either pretreated with 5 μM BI-605906 for 30 min or left untreated. Then the cells were either stimulated with 10 ng/ml TNF-α or left unstimulated for 15 min. Finally, the cells were lysed and Western-blotting was performed to examine the phosphorylation of SRC-3 at S857, HSP27 at S82 and expression of total SRC-3, HSP27, IKK-α and IκBα using anti-P-S857-SRC-3, anti-P-HSP27, anti-SRC-3, anti-HSP27, anti-IKK-α and anti-IκBα antibodies respectively. The full-length blots are presented in supplementary figure S34.