Journal: bioRxiv

Article Title: LncRNA YIYA enhances pancreatic cancer proliferation under high-glucose conditions through RAS–PKM2–mediated metabolic reprogramming that reinforces the Warburg phenotype

doi: 10.1101/2025.10.30.685546

Figure Lengend Snippet: (a) Relative change in Oxygen consumption rate (OCR) post-YIYA knockdown in MIA PaCa-2 cells grown under HG conditions. (b) Relative change in the ATP levels post-YIYA knockdown in MIA PaCa-2 cells grown under HG conditions. (c) Relative mRNA expression of GLUT1 upon YIYA inhibition in MIA PaCa-2 cells grown under HG conditions. (d) Relative fold change in glucose uptake post-YIYA inhibition in MIA PaCa-2 cells grown under HG conditions. (e) Immunofluorescence images representing PKM2 expression and localization in MIA PaCa-2 cells grown under NG or HG conditions (Scale Bar: 20 µm). (f) Immunoblot images representing PKM2 expression in MIA PaCa-2 cells grown under NG or HG. (g) Relative mRNA expression of LDH in MIA PaCa-2 cells grown under NG or HG. (h) Relative mRNA expression of MPC in MIA PaCa-2 cells grown under NG or HG. (i) Relative mRNA expression of PKM2 in HG cultured MIA PaCa-2 cells upon YIYA inhibition. (j) Immunoblot images representing PKM2 expression in MIA PaCa-2 cells grown under HG upon YIYA inhibition. (k) Relative fold change in extracellular lactate concentration in HG cultured MIA PaCa-2 cells upon YIYA inhibition. The glucose concentration used for NG is 5.5 mM, and for HG is 25 mM. The final concentration of siRNA used for transfection was 40 nM. Unless otherwise specified, treatments were carried out for 48 h. For quantitative comparisons, bar graphs representing gene expression or protein levels were normalized to the respective housekeeping gene or loading control (either GAPDH or β-actin). Fold change values shown in the graphs are expressed relative to the control, which was set to 1. All results are presented as mean ± SD from at least three independent experiments (n = 3). For comparisons involving multiple groups across varying concentrations and/or time points, two-way ANOVA followed by Tukey’s multiple comparisons post-test was performed. Statistical significance is indicated as (*) p < 0.05, (**) p<0.01, and (***) p < 0.001 versus the control. For pairwise comparisons between two groups, unpaired t-tests were applied.

Article Snippet: KRAS inhibitor (HY-130149), AKT inhibitor (HY-10115), Protein A/G magnetic beads (HY-K0202), and PKM2 inhibitor (HY-103617) were procured from MedChemExpress.

Techniques: Knockdown, Expressing, Inhibition, Immunofluorescence, Western Blot, Cell Culture, Concentration Assay, Transfection, Gene Expression, Control

Journal: bioRxiv

Article Title: LncRNA YIYA enhances pancreatic cancer proliferation under high-glucose conditions through RAS–PKM2–mediated metabolic reprogramming that reinforces the Warburg phenotype

doi: 10.1101/2025.10.30.685546

Figure Lengend Snippet: (a) Relative mRNA and (b) protein expression of KRAS upon YIYA inhibition in MIA PaCa-2 cells grown under HG conditions. (c) Interaction of lncRNA YIYA with KRAS protein in MIA PaCa-2 cells grown under HG conditions, as analyzed using RIP-qPCR (KRAS was immunoprecipitated using a specific antibody, and the associated RNA was analyzed by qPCR). IgG served as a control. RIP-qPCR illustration prepared with Biorender software. (d) Change in mRNA expression of PKM2 upon KRAS inhibition (KRASi; Adagrasib dose∼100 nM) in MIA PaCa-2 cells grown under HG conditions. (e) Immunoblot images representing PKM2 protein expression in MIA PaCa-2 cells grown under HG upon KRAS inhibition (Adagrasib dose∼100 nM). (f) Interaction of PKM2 protein with KRAS protein in MIA PaCa-2 cells grown under NG or HG conditions, as analyzed using CO-IP (KRAS was immunoprecipitated using a specific antibody, and the precipitated complexes were analyzed by SDS–PAGE followed by immunoblotting with a PKM2 antibody.) (g) Interaction of lncRNA YIYA with PKM2 protein in MIA PaCa-2 cells grown under HG conditions, as analyzed using RIP-qPCR (PKM2 was immunoprecipitated using a specific antibody, and the associated RNA was analyzed by qPCR). IgG served as a control. (h) Immunoblot images representing KRAS protein expression in MIA PaCa-2 cells grown under HG upon YIYA inhibition and autophagy inhibition (CQ∼10 μM). The glucose concentration used for NG is 5.5 mM, and for HG is 25 mM. The final concentration of siRNA used for transfection was 40 nM. Unless otherwise specified, treatments were carried out for 48 h. For quantitative comparisons, bar graphs representing gene expression or protein levels were normalized to the respective housekeeping gene or loading control (either GAPDH or β-actin). Fold change values shown in the graphs are expressed relative to the control, which was set to 1. All results are presented as mean ± SD from at least three independent experiments (n = 3). For comparisons involving multiple groups across varying concentrations and/or time points, two-way ANOVA followed by Tukey’s multiple comparisons post-test was performed. Statistical significance is indicated as (*) p < 0.05, (**) p<0.01, and (***) p < 0.001 versus the control. For pairwise comparisons between two groups, unpaired t-tests were applied.

Article Snippet: KRAS inhibitor (HY-130149), AKT inhibitor (HY-10115), Protein A/G magnetic beads (HY-K0202), and PKM2 inhibitor (HY-103617) were procured from MedChemExpress.

Techniques: Expressing, Inhibition, Immunoprecipitation, Control, Software, Western Blot, Co-Immunoprecipitation Assay, SDS Page, Concentration Assay, Transfection, Gene Expression