glycolysis antibody sampler kit  (Cell Signaling Technology Inc)

Journal: iScience

Article Title: CCL20-CCR6 signaling alters the metabolic reprogramming to promote the pathogenic Th17 cell differentiation

doi: 10.1016/j.isci.2025.114385

Figure Lengend Snippet: CCR6 signaling alters the energy metabolism in Th17 cells (A) Mouse naive T cells (CD4 + CD25 − CD44 lo ) were in vitro differentiated into pathogenic and non-pathogenic Th17 cells. Transcriptomic analysis of Th17 cells cultured in the presence or absence of CCL20 was performed. A heatmap of the fold change (log2) for differentially expressed genes that participate in glycolysis and oxidative phosphorylation is shown. Each line represents one gene, and each column represents one sample. Different colors represent the expression levels (from red to green: increased expression). (B) GSEA enrichment plots of gene sets in Th17 + CCL20 cells compared with Th17 cells, and showing the enrichment scores of oxidative phosphorylation. (C) Glucose uptake was monitored using 2-NBDG by using Flow cytometry and shown as a stacked histogram (left), and the MFI of 2-NBDG uptake (right). (D) After four days of culture of naive CD4 T cells, lactate secretion in the cultured media was measured. The error bar represents ±SD. Experiments were performed with four technical replicates. (E) The extracellular acidification rate (ECAR) and (F) the oxygen consumption rate (OCR) were measured. Error bar represents ± S.E.M. Data shown represents one of the three independent experiments. One-way ANOVA followed by Tukey’s test was used. ∗ p < 0.01, ∗∗ p < 0.001, ∗∗∗ p < 0.0001, ∗∗∗∗ p < 0.0001. (G) The expression of enzymes involved in glycolysis, and (H) a few of the enzymes involved in the TCA cycle in Th17 (non-pathogenic) and Th17 + CCL20 (pathogenic) cells were monitored using Western blot. The data shown represents one of the two independent experiments.

Article Snippet: Glycolysis Antibody Sampler Kit , Cell Signaling Technology , Cat# 8337T; RRID: AB_10897509.

Techniques: In Vitro, Cell Culture, Phospho-proteomics, Expressing, Flow Cytometry, Western Blot

Journal: Scientific reports

Article Title: Exercise promotes peripheral glycolysis in skeletal muscle through miR-204 induction via the HIF-1α pathway.

doi: 10.1038/s41598-025-85174-0

Figure Lengend Snippet: Fig. 2. Chronic exercise mouse model decreases serum insulin level, but increases glucose clearance and glycolysis. (A) miR-204 expression levels in serum of chronic exercise Con (control) and treadmill (TM) groups. Rplp0 was used for an internal control. (B) miR-204 chronic exercise Con and TM groups. Rplp0 was used for an internal control. (C) Blood glucose (mg/dl) and serum insulin levels (uIU/ml) of chronic exercise Con and TM groups. (D) Glucose tolerance test (GTT) of chronic exercise Con and TM groups. (E) Western blot analysis and quantification of glycolytic enzymes (HK2; hexokinase 2, PKM2; pyruvate kinase M2, PDH; pyruvate dehydrogenase, LDH; lactate dehydrogenase) in skeletal muscles (TA; tibialis anterior muscle, GAS; gastrocnemius, QF; quadriceps femoris) of Con and TM groups. HSP90 was used for an internal control. Number of mice used for experiment: Chronic exercise Con (6), Routinely exercised TM (6).

Article Snippet: Primary antibodies used: HK2, PKM2, PDH, LDH (#8337, Cell Signaling Technology, Danvers, United States), HSP90 (#4874, Cell Signaling Technology), HIF1α (A7684, Abclonal), SREBP-1 (sc-13551, Santa Cruz Biotechnology, CA, United States), CD36 (A5792, Abclonal, Woburn, Unisted States), pACC (#3661, Cell Signaling Technology), ACC (#3676, Cell Signaling Technology).

Techniques: Expressing, Control, Western Blot, Muscles

Journal: Scientific reports

Article Title: Exercise promotes peripheral glycolysis in skeletal muscle through miR-204 induction via the HIF-1α pathway.

doi: 10.1038/s41598-025-85174-0

Figure Lengend Snippet: Fig. 5. In vitro miR-204 mimic increases myoblast glycolysis. (A) miR-204 expression levels of C2C12 cells transfected with PBS (Con) and miR-204 mimic. Rplp0 was used for an internal control. (B) mRNA expression levels of glycolysis enzymes (Hk1; hexokinase1, Hk2; hexokinase2, Pfk2; phosphofructokinase2, Pfkm; phosphofructokinase M, Pkm; pyruvate kinase M) in C2C12 cells transfected with PBS and miR-204 mimic. Rplp0 was used for an internal control. (C) Extracellular acidification rate (ECAR) of Con and miR-204 mimic groups. ECAR was normalized by each baseline of well. Glucose was injected to induce glycolysis. Glycolysis was calculated by induced ECAR after glucose injection. (D) Western blot analysis and quantification of glycolytic enzymes (HK2; hexokinase 2, PKM2; pyruvate kinase M2, PDH; pyruvate dehydrogenase, LDH; lactate dehydrogenase) in Con and miR-204 mimic groups. HSP90 was used for an internal control. (E) Oxygen consumption rate (OCR) of Con and miR-204 mimic groups. OCR was normalized by each baseline of well. 1.5 µM of Oligomycin, 0.5 µM of carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP), and 0.5 µM of rotenone/antimycin (rot/ant) were injected. ATP production rate was calculated by decreased OCR after oligomycin treatment. Maximal respiration rate was calculated by decreased OCR after rot/ant treatment. All experiments were performed at least three times.

Article Snippet: Primary antibodies used: HK2, PKM2, PDH, LDH (#8337, Cell Signaling Technology, Danvers, United States), HSP90 (#4874, Cell Signaling Technology), HIF1α (A7684, Abclonal), SREBP-1 (sc-13551, Santa Cruz Biotechnology, CA, United States), CD36 (A5792, Abclonal, Woburn, Unisted States), pACC (#3661, Cell Signaling Technology), ACC (#3676, Cell Signaling Technology).

Techniques: In Vitro, Expressing, Transfection, Control, Injection, Western Blot