Fig. 3: Acetylation affected phosphorylation of SRPK1 and SRSFs in cisplatin-resistant cells.

a MCF7 cells were transfected with Flag-tagged Tip60 (wt) or HAT-deficient mutant (mut). The PTMs of SRPK1 were assessed by immunoprecipitation with Ac-K and p-Ser antibodies. b MCF7 cells were transfected with Myc-tagged SRPK1 (wt) or Mut7. The phosphorylation of SRSFs was evaluated by using the mAb104 antibody. c MCF7 and MCF7R cells were treated with the indicated concentrations of cisplatin for 5 days and lysed for immunoblotting for SRPK1 (K1) using the Phos-tag gel. d In cisplatin-treated MCF7 and MCF7R cells, SRSF phosphorylation was detected by mAb104. e MCF7R cells were transfected with SRPK1 siRNA (siK1) with or without concurrent overexpression of Flag-tagged Tip60. The effect on SRSF phosphorylation was examined by mAb104. f 231 and 231R cells were treated with DMF or cisplatin. Then the relative levels of SRPK1 acetylation and phosphorylation were determined by using the Ac-K and p-Ser antibodies in immunoprecipitation. g 231 cells were transfected with Myc-tagged SRPK1 (wt) or Mut7. The immunoprecipitation was performed with the phospho-Ser beads, followed by immunoblotting with the c-Myc antibody. h In cisplatin-treated 231 and 231R cells, the phosphorylation of SRSFs was checked by mAb104. i Flag-tagged Tip60 (wt) or HAT-deficient mutant (mut) was introduced into 231R cells. Immunoblotting was then performed with mAb104. j Flag-tagged Tip60 constructs were co-expressed with Myc-tagged SRPK1 constructs as indicated. The SRSF phosphorylation level was evaluated with mAb104. For Phos-tag gel in (c) and Western blots in the other panels, they are representative of three experiments with similar results.