Fig. 4: TM4SF5-mediated CD44v8-10 expression increased CD98hc enrichment at the plasma membrane for intracellular GSH-mediated ROS modulation.

a Cells were transfected with control, CD44s, or CD44v8-10 plasmids, and intracellular GSH levels were measured. b, c Cells were transduced with the indicated retrovirus and/or lentivirus to target certain sequences of molecules (Table 1) and intracellular GSH levels were measured. Another set of cells was also processed for immunoblot analysis (bottom in c). d–f Cells transduced with different retrovirus or lentivirus without (d, f) or with erastin treatment (e) were processed for cystine uptake analysis. g Cells stably transduced with control (−) or shTM4SF5 (#2 or #4 sequence, Table 1) lentivirus were treated without or with cystine in glutamate-free culture media before glutamate secretion measurement. h–j Cells stably transduced with HA control (−) or HA-TM4SF5 retrovirus (h), transfected with siRNA for control (NS) or siCD44v8-10 sequence (#1 or #2, Table 1, i), or transfected with FLAG-TM4SF5 and HA-xCT plasmids, were treated without (i) or with vehicle (control) or TNF-α (h, j). Quantitative immunofluorescent images were analyzed for CD98hc (h) or xCT (j) localization at the plasma membrane or Pearson’s values for co-localization between TM4SF5 and CD98hc at the plasma membrane (i). The p values were analyzed by ANOVA with Tukey’s range-test. P values <0.05 were considered statistically significant. Data represent three isolated experiments. See also Figures S4 and S5