Fig. 4: PKLR rescues ZBTB10-decreased glucose metabolism. | Cell Death & Disease

Fig. 4: PKLR rescues ZBTB10-decreased glucose metabolism.

From: Pyruvate kinase L/R links metabolism dysfunction to neuroendocrine differentiation of prostate cancer by ZBTB10 deficiency

Fig. 4

A–C Quantification of glucose uptake (A), lactate amounts (B), and pyruvate levels (C) by colorimetric assays of LNCaP and C4-2 cells expressing the non-target control (NC) or PKLR shRNA vector and incubated with charcoal-stripped serum (CSS)-containing medium for 48 h. n = 3 per group. * vs. the NC; # vs. -CSS; by a two-way ANOVA. D Relative PKLR mRNA levels in LNCaP and C4-2 cells stably expressing the NC or PKLR shRNA vector. n = 3 per group. * vs. the NC; # vs. -CSS. **p < 0.01, ***p < 0.001; by a two-way ANOVA. E-G Quantification of glucose uptake (E), lactate amounts (F), and pyruvate levels (G) by colorimetric assays of LNCaP and C4-2 cells expressing the empty vector (EV) or ZBTB10 cDNA vector and incubated with CSS-containing medium for 48 h. n = 3 per group. * vs. the EV; # vs. -CSS; by a two-way ANOVA. Relative glucose uptake, lactate amounts, and pyruvate levels are presented as the mean ± SEM from three biological replicates. *p < 0.05, **p < 0.01, ***p < 0.001. H Relative PKLR mRNA levels in LNCaP and C4-2 cells stably expressing the EV or ZBTB10 cDNA vector. n = 3 per group. * vs. the EV; # vs. -CSS. *p < 0.05, **p < 0.01; by a two-way ANOVA. I, J Bioenergetics trace from the Seahorse analysis showing values of the extracellular acidification rate (ECAR) in C4-2 cells with expression of the NC or PKLR shRNA vector (I) and EV or ZBTB10 cDNA vector (J) following treatment with CSS-containing medium for 48 h, and incubated with 12 mM d-glucose and 50 mM 2-deoxyglucose (2-DG). * vs. -CSS; # vs. the NC (I) or EV (J); by a two-way ANOVA. K PKLR and ZBTB10 protein levels of C4-2 and LNCaP cells stably express the EV, ZBTB10, or ZBTB10+PKLR cDNA vector. L Relative PKLR, ZBTB10, NE marker, stem cell marker, and androgen-responsive gene mRNA levels in C4-2 cells expressing the EV, ZBTB10, or ZBTB10 + PKLR cDNA vector. * vs. the EV; # vs. ZBTB10; by a two-way ANOVA. Quantification of mRNA is presented as the mean ± SEM from three biological replicates. *p < 0.05, **p < 0.01, ***p < 0.001. M Bioenergetics trace from the Seahorse analysis showing ECAR values in C4-2 cells with expression of the EV, ZBTB10, or ZBTB10+PKLR cDNA vector, and incubated with 12 mM d-glucose and 50 mM 2-DG. N Bioenergetics trace from the Seahorse analysis showing oxygen consumption rate (OCR) values in C4-2 cells with stable EV, PKLR, or PKLR+ZBTB10 cDNA vector expression, and incubated with 1 μM oligomycin, 0.75 μM FCCP, and 0.5 μM each of rotenone and antimycin A. * vs. the EV; # vs. ZBTB10; by a two-way ANOVA. Relative ECAR and OCR values are presented as the mean ± SEM from three biological replicates. *p < 0.05, **p < 0.01, ***p < 0.001.

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