Fig. 6: Interaction between glycolysis and mTOR controls the proliferation of ALDH-high endometrial cancer cells. See also Supplementary Figs. S9, S10, and S11.

A In silico screening of RARA target genes connected to mTORC1 signaling using a combination of public chromatin immunoprecipitation (ChIP) database and gene set enrichment analysis based on microarray analysis of endometrial cancer spheroid cells. Combined with the 85 genes from 184 genes in the gene set of hallmark of mTORC1 signaling and putative RARA-regulated genes retrieved from ChIP-Atlas (http://chip-atlas.org/), eight genes were identified as the candidate genes correlated to ALDH-RARA-mTOR axis. Western blot analyses of B ALDH-high and ALDH-low spheroids cells, C the infected spheroids cells, and D disulfiram treatment. E Relative LDHA activity of control bulk, ALDH-high, and exogenous ALDH1A1 overexpressing spheroid cells. F Volumes (mean ± SEM) of xenograft tumors from 1 × 10⁵ EMN24 spheroid cells with the presence or absence of AZ-33 in vivo treatment. n = 6, Student’s t-tests. Xenograft tumor images post-excision on day 27 are on the right. Scale bar: 10 mm. G Relative ALDH-high and ALDH-low spheroid cell viability with the indicated in vitro AZ-33 treatment for 4 days. H Western blot analyses of spheroids cells after AZ-33 treatment for 24 h. I Relative spheroid cell viability in the presence or absence of 100 µM AZ-33 and/or 1 µM MHY1485 in vitro treatment for 4 days. J Western blot analyses of spheroids cells after everolimus or MHY1485 treatment for 24 h. K Relative glucose uptake of spheroid cells with 80 µM everolimus in vitro treatment. L Relative glycolytic rate of spheroid cells with 25 µM everolimus or 10 mM 2-DG in vitro treatment.