Fig. 2: FGFR2-mediated necroptosis is RIP1 and MLKL-dependent.

a KYSE140 and KYSE180 cells were treated with 40 μM AZD4547 in the absence or presence of 30 μM Nec-1, 5 μM NSA, 20 μM ZVAD and 1 μM Fer-1 for 24 h to assay cell viability. b The release of LDH was detected following AZD4547 with Nec-1 and NSA for 24 h. c, d Knockdown of RIP1 and MLKL inhibited AZD4547-induced cell death (c) and LDH release (d). The p value was derived from a comparison with the control group. e–g Cells were transfected with FGFR2 (e), FGFR1 (f) or FGFR3 (g) siRNAs, and whole-cell lysates were subjected to western blot analysis after 48 h. pMLKL was induced by adding 20 μM AZD4547. h Cells were analyzed for proliferation by a real-time cell analyzer (RTCA)-MP system. i Knockdown of FGFR2 inhibited 10 μg/mL cisplatin-induced LDH release. j Transmission electron microscopy of the KYSE140 and KYSE180 cells transfected with FGFR2 siRNAs for 48 h. Yellow arrowheads denote loosened nuclei. Red arrowheads indicate swelled cellular organelles. Blue arrowheads indicate plasma membrane rupture. Green arrowheads indicate vacuolization. Scale bar: 2 μm. k After 48 h transfection with FGFR2 siRNA, membrane-cytosolic extraction proteins were measured using western blot. ns not significant. All data are mean ± SD from n = 3 (b, d), 4 (c, i) or 5 (a) biological replicates. Statistical analyses were performed using one-way ANOVA with multiple comparisons. Source data are provided as a Source Data file.