Fig 1: Effects of VT on the proteins involved in glucose and lipid metabolism. The expression of phosphorylated protein kinase B (p-Akt), Akt, phospho-Forkhead box protein O1 (p-Foxo1), Foxo1, sterol response element-binding protein 1c (SREBP1c), fatty acid synthase (Fasn), and phosphoenolpyruvate carboxykinase 2 (Pck2) was detected by WB, and Gapdh was simultaneously detected as a standard internal reference protein (A); the expression ratio of p-Akt to Akt in each group (B); the expression ratio of p-Foxo1 to Foxo1 in different group (C); the protein expression of SREBP1c in four groups (D); the protein expression of Fasn in each group (E); the protein expression of Pck2 in all groups (F); and the protein expression of Foxo1 in the respective groups (G). The data are presented as floating bars (min-max, three repetitions), and the CON group was used for standardization. * p < 0.05, *** p < 0.001 contrasted with the CON group, # p < 0.05 compared to the MOD group, and p < 0.05 compared with the MET group, n = 6.
Fig 2: A Summary of the Observed Metabolic States of Quiescent, LPS Stimulated, and COPD NeutrophilsA diagram showing the metabolic states of resting (A), stimulated (B), and COPD (C) neutrophils showing increased glycolytic activity and glycogen synthesis in response to LPS and defective glycogen cycling and glycolysis in COPD. Genes identified to actively regulate neutrophil glucose transport (Glut1), gluconeogenesis (GNG) (Fbp1 and Pck2), glycogenesis (Gys1, Gbe1, and Ugp2), and glycogenolysis (Pygl) are highlighted in red. Arrow thickness indicates the relative flux through metabolic pathways with glycogenolysis and glucose oxidation highlighted in blue and glycogenesis and gluconeogenesis in green.
Fig 3: PEPCK-M is essential for Ser/Gly synthesis. a, b 13C enrichment of serine and glycine in HeLa cells exposed to 2 mM [U-13C]glutamine for 4 h in the DMEM media containing 10% dFCS and 0 mM glucose. Incorporation of 13C was analyzed using GC-MS. Statistical significance compared with shCtrl was determined by unpaired two-tailed Student’s t test. c Quantitative real-time PCR analysis of PCK2 mRNA expression levels in shCtrl HeLa cells grown in 0 mM glucose MEM media with or without 0.4 mM serine and 0.4 mM glycine (Ser/Gly) for 6 or 12 h. Values were normalized to time 0 h. Two-way ANOVA with Sidak multiple comparison post-test analysis indicate significance of Ser/Gly effect (*). d Cell viability was measured in HeLa cells using an MTT assay after 72 h of growth in 0 mM glucose MEM media with or without 0.4 mM serine and 0.4 mM glycine (Ser/Gly). The sensitivity of each cell line to the addition of serine and glycine is shown as fold change with respect to the viability in the presence of +Ser/Gly. Two-way ANOVA with Sidak multiple comparison post-test analysis indicate significance vs + Ser/Gly (*)
Fig 4: PCK2 promotes inflammatory responses in Kupffer cells. (A) The enzyme activity of PCK2 was measured after 3MP treatment in Kupffer cells. (B) The mRNA expression of cytokines including IL-6, MCP1, and IL-10 was measured by real-time PCR. (C) The production of cytokines in the medium was measured by cytometric bead array (CBA) after 3MP treatment. (D) The protein expression of PCK2 was determined by Western blot after siRNA knockdown. (E) The mRNA expression of IL-6 and IL-10 was measured by real-time PCR after PCK2 knockdown. (F) The production of cytokines was determined by CBA after PCK2 knockdown. (G) The protein expression of PCK2 was determined by Western blot after vector transfection. (H) The mRNA expression of cytokines including IL-1β, IL-6, TNFα, and IL-10 was measured by real-time PCR. (I) The production of cytokines was determined by CBA after PCK2 overexpression (PCK2 OE). Values are mean ± SEM. *p < 0.05, **p < 0.01. values with different superscript letters mean significant differences.
Fig 5: PCK2 is increased in Kupffer cells after acute LPS administration in mice. (A) Immunohistochemical staining and quantification of F4/80 and CD68 in the liver (magnification, x400). (B) The expression levels of inflammatory cytokines were determined by real-time PCR. (C) The levels of inflammatory cytokines in plasma were determined by cytometric bead array (CBA). (D) Co-immunofluorescence staining of PCK2 and CD68. Values are mean ± SEM, n = 6. **p < 0.01 versus control group.
Supplier Page from Abcam for Anti-PCK2 antibody