PKM2 Antibody - #AF5234

Fig. 4. | Bruceine A induces cell growth inhibition and apoptosis via PFKFB4-mediated glycolysis in MIA PaCa-2 cells. (D) MIA PaCa-2 cells were treated with 25 nM bruceine A for 0, 6, 12, and 24 h. Immunoblots against GLUT1, HK2, PFKFB4,PFKM, PKM2, LDHA, and β-actin from cell lysates of MIA PaCa-2 cells were detected. β-actin was served as control. Results were expressed as means ± SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 versus control cultured with 0.1% DMSO by one-way ANOVA and post hoc tests.

Fig. 6 The relationship between PTP1B and AMPK. a PTP1B overexpression resulted in decreased p-AMPK (alpha). b, c The negative correlation between PTP1B and p-AMPKα was showed in pancreatic cancer patient tissue samples (p < 0.001, p value was obtained by a Pearson χ2 test; scale bar, 200 μm and 50 μm). d PTP1B inhibition either by shRNAs or by LXQ46 increased the phosphorylation of PKM2. e, f The inactivated PKM2 resulted in increased phosphorylation of AMPKα and decreased the phosphorylation of PRAS40, causing the inhibition of mTOC1 activity. g PTP1B inhibition caused AMPK activation and decreased p-p70S6K in vivo (scale bar, 200 and 50 μm).

Fig. 5. Pimozide promotes the expression of p53 through PI3K/Akt/MDM2 pathway. (A-B) Cells were treated with the indicated concentrations of Pimozide for 24 h, the protein expression of p-PI3K, PI3K, p-Akt, Akt, p-MDM2, MDM2 in MCF-7 (A) and MDA-MB-231 (B) cells were determined by Western blot analysis (left panel). Densitometry analysis was performed to assess the protein expression of p-PI3K/PI3K, p-Akt/Akt, p-MDM2/MDM2 (normalized to β-actin expression) (right panel). (C-F) Western blot analysis for p-PI3K, PI3K, p-Akt, Akt, p-MDM2, MDM2, P53, and PKM2 protein levels in MCF-7 (C, E) and MDA-MB-231 (D, F) cells incubated with PI3K inhibitor LY294002 or PI3K agonist SC79 as indicated. (G) After treatment with SC79, colony-forming assay images (left panel) and quantification of colony number percentages (right panel). Data represent mean ± SD from three biological replicates (*p < 0.05, **p < 0.01, ns: not significant).

Fig. 3 NCAPD3 induces aerobic glycolysis in LC cells. (A) Single-gene GSEA analysis of NCAPD3. (B) Glucose uptake in NCAPD3-knockdown H1975 cells and NCAPD3-over-expression HCC827 cells was measured with flow cytometry by using the fluorescent glucose analog 2-NBDG. (C) Lactate production was measured in LC cells with NCAPD3-knockdown or NCAPD3-over-expression using the Lactic Acid (LA) Content Assay Kit. (D) The extracellular acidification rate (ECAR) of NCAPD3-knockdown H1975 cells and NCAPD3-over-expression HCC827 cells was also monitored. (E) The ATP levels in NCAPD3-knockdown H1975 cells and NCAPD3-over-expression HCC827 cells were quantified using the ATP Assay Kit. (F) The protein levels of NCAPD3 and aerobic glycolysis-related targets HK2, PKM2, GLUT1, and LDHA were determined by Western blotting. ▲P