Figure 4.

TNIK binds to the ECD domain to phosphorylate EGFR and regulate ferroptosis resistance

(A) Immunoprecipitation of TNIK in C4-2 cells followed by immunoblot analysis of EGFR or TNIK.

(B) Immunoprecipitation of EGFR in C4-2 cells followed by immunoblot analysis of EGFR or TNIK.

(C) Co-immunofluorescence (co-IF) analysis of TNIK and EGFR proteins in C4-2 cells after transfected TNIK wild-type vector. Scale bars: 50 μm.

(D) Schematic diagram of the Myc-EGFR constructs. C4-2 cells were transfected with the indicated plasmids for 48 h. The cell lysates were immunoprecipitated with anti-TNIK immunomagnetic beads and immunoblotted with the indicated antibodies.

(E) The levels of EGFR/p-EGFR and EGFR-related protein were measured in C4-2 cells after transfected TNIK wild-type vector.

(F) The levels of EGFR/p-EGFR and EGFR-related protein were measured in C4-2 cells after transfected siTNIK.

(G) Expression of a TNIK kinase mutant (D171A) reduces phosphorylation of EGFR.

(H) The levels of p-EGFR and NRF2 protein were measured in C4-2 cells after knocking down the ECD domain.

(I) Levels of cell death in C4-2 cells treated with 4 μm erastin (up). Levels of lipid peroxidation in C4-2 cells following the same treatment (down). Data are shown as mean ± SD. ∗∗∗p < 0.005.