Fig. 7: YAP m⁵C modification and downstream exosome secretion-promoting transcription system causes AZD9291 resistance.

A–D IC50 was calculated in AZD9291 (24 h) treated H1975 AZD9291 resistant (H1975^Res, A), normal H1975 (B), HCC827^Res (C), normal HCC827 (D) cells with indicated treatment. GW4869 (10 µM, 24 h). E 3D spheroids were generated by NSUN2 with or without ALYREF knocked out H1975^Res and HCC827^Res cells. After the formation of 3D spheroids, AZD9291 (10 µM, 24 h) with or without GW4869 (10 µM, 24 h) was treated. Then cells were stained with SYTOX green. Representative images and graphed data are shown, Scale bar, 50 µm. F 3D spheroids were generated by YAP with or without Mycn and SOX10 overexpressed H1975 and HCC827 cells. After the formation of 3D spheroids, AZD9291 (10 µM, 24 h) was treated. Then cells were stained with SYTOX green. Representative images and graphed data are shown, Scale bar, 50 µm. G–I Indicated protein levels (G), YAP m⁵C level (H), exosome concentration (I) were measured in H1975, H1975^Res, HCC827 and HCC827^Res cells. The level of proteins was normalized to that of GAPDH, and the normalized level of proteins in negative control groups were arbitrarily set to 1. The data are shown as the means ± SD from 3 to 5 independent experiments. Images of IB are representative ones of three independent experiments. ^*p < 0.05, ^**p < 0.01 indicate statistical significance. The data from (A–D) were analyzed by a two-way ANOVA test. The data from (H, I) were analyzed by a student’s t test.