Fig 1: PCK2 promotes glycolysis via regulating PFKP in vitro. (A) Scatter plot (A), the color of wathet represents PFKP, the color of navy blue represents PFKM, the color of green represents PFKLshowed that the expression of the three isoforms of PFK was upregulated in PCK2 overexpression group in GM60, compared to vector group. (B-D) qPCR analysis showed that expression of PFKP, PFKM, PFKL in the PCK2 overexpression group was upregulated, compared with vector group in 3D ECM with tunable stiffness (*P < 0.05, ***P < 0.001, compared to vector cells, #P < 0.05, ##P < 0.01, ###P < 0.001, ####P < 0.0001, compared to NC group, by students' t-test). (E, F) Representative images of immunohistochemical staining of PCK2 (green), PFKP (purplish red), and nucleus (blue) of PCK2-overexpression-, PCK2-knockdown-, vector- and NC-transfected BMMSCs cultured in 3D ECM with different stiffness. Scale bar is 50 µm. All the results were repeated three times and represents means ± SEM.
Fig 2: PFKP shows a high expression in IDH-mutant ICCs. (A) Representative images of PFKP-staining of human ICC surgical specimens. Scale bars, 500 µm. (B) PFKP protein expression in human ICC surgical specimens and association with IDH mutation status.
Fig 3: Upregulation of PFKP expression in NSCLC tissue and cells. (A) PFKP expression in NSCLC tissues and normal tissues was analyzed using data from TCGA. (B) Survival analysis based on TCGA data. (C) RT-qPCR was used to detect the mRNA expression levels of PFKP in NSCLC and corresponding adjacent tissues (normal) (n=84). (D) Immunohistochemical analysis was used to detect PFKP expression in NSCLC and corresponding adjacent tissues (normal) (n=84); scale bar, 200 µm. **P<0.01 vs. normal tissues. (E) RT-qPCR was used to detect the mRNA expression levels of PFKP in 16HBE cells and NSCLC cells (H1299, H23 and A549). (F) Western blotting was used to detect the protein expression levels of PFKP in 16HBE cells and NSCLC cells (H1299, H23 and A549). Data are presented as the mean ± SD from three independent experiments. *P<0.05, **P<0.01 vs. 16HBE cells. PFKP, platelet isoform of phosphofructokinase; NSCLC, non-small cell lung cancer; TCGA, The Cancer Genome Atlas; RT-qPCR, reverse transcription-quantitative PCR.
Fig 4: PFKP inhibits apoptosis of non-small cell lung cancer cells. (A) Apoptosis of H23 and H1299 cells was detected by flow cytometry. (B) Expression levels of apoptosis-related proteins (caspase-3 and Bcl-2) were detected by western blot analysis. Data are presented as mean ± SD from three independent experiments. **P<0.01 vs. control group; ##P<0.01 vs. si-NC group; $$P<0.01 vs. pCMV6-ENTRY group. PFKP, platelet isoform of phosphofructokinase; Bcl-2, B-cell lymphoma 2; si, small interfering RNA; NC, negative control.
Fig 5: PFKP promotes the proliferation of non-small cell lung cancer cells. (A) mRNA expression levels of PFKP in H1299 and H23 cells were determined by reverse transcription-quantitative PCR. (B) Viability of H1299 and H23 cells was detected by Cell Counting Kit-8 assay. (C) Colony formation assay was used to detect the proliferation of H1299 and H23 cells. Data are presented as the mean ± SD from three independent experiments. **P<0.01 vs. control group; ##P<0.01 vs. si-NC group; &&P<0.01 vs. si1-PFKP group; $$P<0.01 vs. pCMV6-ENTRY group. PFKP, platelet isoform of phosphofructokinase; OD, optical density; si, small interfering RNA; NC, negative control.
Supplier Page from Abcam for Anti-PFKP antibody [OTI1D6]