Fig 2: GLS expression has a negative spatial correlation with EZH2 in colorectal cancer tissues.A, B Immunofluorescence of EZH2 (A) and GLS (B) in human colorectal cancer samples. DAPI is used to indicate the nucleus.

Fig 3: The EZH2-GLS axis regulates glucose-deprivation-induced cell death by modulating intracellular GSH levels.A Immunoblotting for cleaved-PARP in HCT116 or SW480 cells cultured with or without glucose deprivation (0 mM, 16 h) in presence or not of 4 µM Fer or 10 µM/20 µM NAC (n = 2 independent experiments). B DCFH staining of HCT116 NC and shEZH2 cells under normal condition or glucose deprivation (0 mM, 10 h). C DCFH staining of HCT116 cells pretreated with DMSO or GSK126 (4 µM, 48 h), followed by glucose deprivation treatment (0 mM, 10 h). D Intracellular GSH content of HCT116 cells pretreated with DMSO or BPTES (10 µM, 48 h), followed by glucose deprivation treatment (0 mM, 10 h). E Intracellular GSH content for neutralizing ROS in HCT116 NC and shEZH2 cells pretreated with DMSO or BPTES (10 µM, 48 h), followed by glucose deprivation treatment (0 mM, 10 h). F Intracellular GSH content of HCT116 or SW480 cells pretreated with DMSO or GSK126 (4 µM, 48 h), followed by glucose deprivation treatment (0 mM, 10 h). G DCFH staining of HCT116 cells pretreated with or without BPTES (10 µM, 24 h) or/and GSK126 (4 µM, 72 h), followed by glucose deprivation treatment (0 mM, 10 h). The experiments in A–C and G were repeated twice independently with similar results. In D–F, data are mean ± s.d., n = 3 independent experiments; P-values were calculated using two-tailed unpaired Student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001; ns, non-significant; scale bars, 200 µm.

Fig 4: Gls1 was upregulated in SS patients and could be induced upon activation. (a) Expression of Gls1 in circulating CD4+ T cells obtained from healthy individuals and SS patients. n > 25. (b) Immunohistochemical staining of CD4-positive cells and Gls1-positive cells in the labial glands of SS patients. (c) qRT–PCR analysis of Gls1 mRNA expression in murine CD4+ T cells simulated with anti-CD3 and anti-CD28 antibodies in a time-dependent manner. (d) qRT–PCR analysis of Gls1 mRNA expression in human purified circulating CD4+ T cells simulated with anti-CD3 and anti-CD28 antibodies in a time-dependent manner. (e) Representative Western blot pictures of Gls1 protein expression in human purified circulating CD4+ T cells simulated with anti-CD3 and anti-CD28 antibodies. (f) Representative Western blot pictures of Gls1 protein expression in murine splenic CD4+ T cells simulated with anti-CD3 and anti-CD28 antibodies. (g) Representative picture of Gls1 expression and location in murine CD4+ T cells under a confocal microscope. Blue light indicates the position of nuclei; green light indicates the position of the cytoskeleton; red light indicates the position of mitochondria; violet light indicates the position and strength of Gls1. Summary data are presented as **p < 0.001 with unpaired two-tailed Student's t-tests.

Fig 5: LAPTM5 enhances the resistance to docetaxel in ER+ BC cells via the activation of glutamine-mediated mTOR signaling. (A-C) Cell viability curves of blank (A), LAPTM5-sh3 (knockdown) (C) and LAPTM5-OE (overexpressing) MCF-7 (B) cells treated with 0, 1, 2, 4, 8, or 16 µM docetaxel for 24 h. (D) Statistical analysis of the half maximal inhibitory concentration (IC50) values of the above three cell lines. **P<0.01, compared to the blank MCF-7 cells. (E) Protein expression and densitometric analysis results of key molecules of glutamine metabolism and mTOR signaling in blank, LAPTM5-sh3 and LAPTM5-OE MCF-7 and T47D cells. (F and G) Relative protein levels of LAPTM5 in the different MCF-7 (F) and T47D (G) cells. **P<0.01 and ***P<0.001, compared with the blank MCF-7 and T47D cells. LAPTM5, lysosomal protein transmembrane 5; ER+ BC, estrogen receptor-positive breast cancer; SLC1A5, sodium-dependent neutral amino acid transporter type 2; GLS1, glutaminase 1; S6K1, ribosomal protein S6 kinase 1; 4EBP1, eukaryotic translation initiation factor 4E (eIF4E)-binding protein 1; MMP9, matrix metallopeptidase 9; nuclear factor ?B, NF?B; p-, phosphorylated.