Fig. 4: SRPK1 rescues the reduction of GSK3β Ser9 phosphorylation under gefitinib treatment, independent of its kinase activity. | Oncogene

Fig. 4: SRPK1 rescues the reduction of GSK3β Ser9 phosphorylation under gefitinib treatment, independent of its kinase activity.

From: Serine-arginine protein kinase 1 (SRPK1) promotes EGFR-TKI resistance by enhancing GSK3β Ser9 autophosphorylation independent of its kinase activity in non-small-cell lung cancer

Fig. 4

A GSEA showing a significant association between gefitinib treatment and GSK3 phosphorylation (BIOCARTA_GSK3_PATHWAY) in the GSE75309 database. B Western blot analysis of GSK3β pSer9 expression in cells treated with gefitinib (0, 1, 5, or 10 µM). C IP analysis of the interactions between SRPK1 and GSK3β examined in NCI-H1975 cells. D Co-IP analysis of the interaction between SRPK1 and GSK3β in 293T cells transfected with HA-Vector, HA-SRPK1, Flag-Vector or Flag-GSK3β, as indicated. E Pulldown assay investigation of the interaction between SRPK1 WT and GSK3β WT in vitro. F Pulldown assay investigation of the interaction between SRPK1 and GSK3β variants. G Isotope radiation assay of GSK3β Ser9 phosphorylation status in SRPK1 and GSK3β variants co-incubated in isotope radiation kinase assay buffer for 30 min at 30 °C. H Kinase activity assay of the in vitro phosphorylation relationship between SRPK1 and GSK3β in E. coli (BL21 DE3) cells transfected with His-SRPK1 WT, His-SRPK1 K109A, GST-GSK3β WT, GST-GSK3β S9A, GST-GSK3β K85A, and GST-GSK3β S9A/K85A. I Kinase activity assay of the in vivo phosphorylation relationship between SRPK1 and GSK3β precipitated from lysates of 293T cells expressing tagged peptides (HA-SRPK1 WT and HA-SRPK1 K109A) using HA antibody.

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