Fig 2: Effects of overexpression of miR-489-3P on PC proliferation and metabolism in vivo. (A) Typical images of nude mice tumors (n = 5), (B) subcutaneous tumor weight, (C) subcutaneous tumor volume, (D) miR-489-3p expression in xenografts by qRT-PCR. (E) Typical IHC staining images of xenografts show Ki-67 and PCNA expression. The bar stands for 50 microns. (F) Typical IHC staining images of xenografts show the expression of metabolic indicators (LDHA, GLUT1, HK2, PKM2). The bar stands for 100 microns. (**P < 0.01, ***P < 0.001).

Fig 3: Identification target genes of lnc-CYB561-5 in NSCLC. (A) The co-expression network. This network consists of lnc-CYB561-5 (green) and its 10 direct neighbours (blue). (B and C) Relative protein expression levels of Bsg, Lbr and Sez6l2 in H1299 cells. (D) The RNA level of Bsg in H1299 cells. ***p< 0.001, NS, p > 0.05 vs. the sh-Ctrl group, n = 5. (E) Western blot analysis to determine the levels of Bsg, Lbr and Sez6l2 on RNA pulldown material. (F) RIP assays were performed to test the interaction of lnc-CYB561-5 and Bsg. Relative quantification of lnc-CYB561-5 and 18S rRNA in RNA-protein complexes immunoprecipitated with IgG or Bsg from whole-cell extracts. (G) Validation of Bsg overexpression efficiency, *p < 0.05, NS, p > 0.05 vs. the control group. (H and I) Western blot determination of Pfk1 and Hk2 in H1299 cells. (I) Measurement of ECAR in H1299 cells. (J) Relative glycolysis rates in H1299 cells. (K) CCK-8 assay analysis of cell proliferation in H1299 cells. (L and M) The migration and invasion of H1299 cells. *p < 0.05, **p < 0.01, ***p < 0.001 vs. the indicated group, n = 5. Data are presented as means ± SEM. Multiple group comparisons were performed using one-way ANOVA followed by Tukey's post hoc test

Fig 4: PGC1α manages the mitochondrial expression of HK2 and glycolysis through HSP70. We mutated serine at 400 sites of HSP70 into lysine (a). The mRNA level of HSP70 was verified by qPCR and expressed as the mean ± SD; n = 3, * p < 0.05 vs. control (b). The effect of the overexpression of HSP70 and HSP70 S400K on the expression level of HK2 mitochondria was detected by a mitochondrial extraction experiment (c). The protein/TOM20 ratio is expressed as the mean ± SD; n = 3, * p < 0.05 vs. control, & p < 0.05 vs. SR-18292 and # p < 0.05 vs. SR-18292+HSP70 (d). Immunoprecipitation was used to detect the binding of HK2 to VDAC1 after SKOV3/CDDP cells were treated and expressed as the mean ± SD; n = 3 (e). Therefore, we speculate that PGC1 regulates the binding of HK2 to VDAC1 through HSP70, and then affects the cisplatin resistance of ovarian cancer (f).

Fig 5: PGC1α regulates the function of HK2 in mitochondria through heat shock protein 70 (HSP70). The protein expressions of HSP100, HSP90, and HSP70 were detected by Western blot in SKOV3/CDDP cells (a). The protein/actin ratio is expressed as the mean ± SD; n = 3, * p < 0.05 vs. control and # p < 0.05 vs. cisplatin (b). The luciferase reporter gene was used to detect whether PGC1α can promote the transcription of HSP70 promoter heat shock factor 1 (HSF1) and expressed as the mean ± SD; n = 3, * p < 0.05 vs. control (c). After PGC1α inhibitor SR-19282 and overexpressed HSP70 at the same time, the mitochondria expressions of HK2 (d) were detected. The protein/ translocase of outer mitochondrial membrane 20 (TOM20) ratio is expressed as the mean ± SD; n = 3, * p < 0.05 vs. control and # p < 0.05 vs. SR-18292 (e). After PGC1α inhibitor SR-19282 and overexpressed HSP70 at the same time, the combination HK2–VDAC1 in SKOV3/CDDP cells was detected (f). After PGC1α inhibitor SR-19282 and overexpressed HSP70 at the same time, the mitochondrial membrane potential (g) and apoptosis (h) in SKOV3/CDDP cells were detected and expressed as the mean ± SD; n = 3.