Extended Data Fig. 9. PCK2-ACSL4(T679) phosphorylation axis promotes ferroptosis in vivo.
a-b, The TRCs and bulk tumor cells from HONE1 cells were implanted subcutaneously into female BALB/c nude mice to construct xenograft growth models and exposed to radiotherapy (8 Gy, arrow) (a) or cisplatin (5 mg/kg, arrow) (b). Tumor volume for each group were shown (n = 5). c, Immunoblots showing the expression of PCK2, pSTAT3 and STAT3 in bulk tumor cells and TRCs from HONE1 cells in indicated conditions. One of three experiments is shown. d, The HONE1 cells with or without fibrin gel were implanted subcutaneously into female BALB/c nude mice to construct xenograft growth models and exposed to cisplatin (arrow). Tumor volumes for each group were shown (n = 5). e, H&E staining and Immunohistochemistry of fibrin showing the presence of fibrin gel in TRC tumors. Images are representative of n = 5 images. f, H&E staining and immunohistochemistry of fibrin, PCK2 and ACSL4 pT679 showing the presence of fibrin gel and the level of PCK2 and ACSL4 pT679 in TRC tumors and bulk tumors treated with cisplatin. Images are representative of n = 5 images. g, Flow cytometric analysis of the proportion of EpCAM+ pSTAT3+ in TRC tumors and bulk tumors treated with cisplatin. n = 5. h-i, The HONE1 cells stably transfected with indicated plasmids (h) or PCK2Tet HONE1 cells with or without fibrin gel (i) were implanted subcutaneously into female BALB/c nude mice to construct xenograft growth models. Indicated groups were exposed to cisplatin (arrow). Mice were provided with a Dox drink (100 mg/kg, every two days, starting from ‘arrowhead’, until the endpoint) or a normal drink. Tumor volumes for each group were shown (n = 5). Data are shown as mean ± SEM (a-b, d, h-i) or mean ± SD (g), unpaired two-tailed t-test (g) or two-way ANOVA (a-b, d, h-i).