cells hk2  (ATCC)

Journal: The Journal of Clinical Endocrinology and Metabolism

Article Title: Propionate Induces Energy Expenditure via Browning in Mesenteric Adipose Tissue

doi: 10.1210/clinem/dgaf280

Figure Lengend Snippet: Effects of propionate on adipocyte browning, glucose uptake, and glycolysis. A to C, Relative messenger RNA (mRNA) expression of UCP1, PGC1α, and GLUT4 in adipocytes in response to propionate (1 mM) treatment vs vehicle control (mean ± SEM, n = 6-9). D, Glucose uptake in adipocytes. Ins + Pro: 1 μM insulin and 1 mM Propionate (mean ± SEM, n = 11-12). Unt: untreated; Ins: 1 μM insulin; Ins + Con:1 μM insulin and vehicle control. E and F, Relative mRNA expression of HK2 and PKM in adipocytes in response to propionate (1 mM) treatment vs vehicle control (mean ± SEM, n = 6-9). # P less than .05, ## P less than .01, #### P less than .0001 propionate vs vehicle control; P less than .05 vs Unt group; * P less than .05, ** P less than .01, **** P less than .0001. Abbreviations: A, adipocyte; M, mesenteric; O, omental; S, subcutaneous;.

Article Snippet: The relative expression levels of mRNA of FFAR2 (Hs00271142 s1), CEBPα (Hs00269972 s1), UCP1 (Hs00222453 m1), PPARγ (Hs01115513 m1), PGC1α (Hs00173304 m1), GLUT4 (Hs00168966 m1), tumor necrosis factor α (TNFα) (Hs00174128 m1), interleukin-6 (IL6) (Hs00174131 m1), FASN (Hs01005622 m1), PKM (Hs00761782 s1), and HK2 (Hs00606086 m1) were compared to the vehicle control and normalized to 18S (Hs03003631 g1), using the 2 −ΔΔCt formula.

Techniques: Expressing, Control

Journal: Clinical and Translational Medicine

Article Title: Exosomal miR‐3126‐5p derived from cancer‐associated fibroblasts facilitates glycolysis to accelerate NSCLC progression by targeting KLF13 to activate the SH2B1/IRS1 axis

doi: 10.1002/ctm2.70554

Figure Lengend Snippet: SH2B1 deficiency refrained glycolysis of NSCLC cells. NSCLC cells were transfected with sh‐SH2B1#2 or SH2B1 overexpression plasmid for 48 h. (A–D) Glucose uptake (A), lactate production (B), ECAR (C), and OCR (D) of NSCLC cells were detected by commercial kits, respectively. (E) Protein abundance of HK2, GLUT1, PDK1, ENO1, and LDHA was assessed by western blotting. (F) GLUT1 expression in NSCLC tissues and paired normal lung tissues was evaluated by immunohistochemical staining. Scale bar = 50 µm. (G) RT‐qPCR analysis of SH2B1 and GLUT1 mRNA levels in NSCLC and normal lung tissues ( n = 40). (H) Correlation between SH2B1 and GLUT1 expression in NSCLC tissues was analyzed by the Pearson correlation analysis ( n = 40). (I) The correlation between SH2B1/GLUT1 expression and survival of NSCLC patients was analyzed by Kaplan‐Meier plotter ( n = 40). Student's t ‐test (for G) or one‐way ANOVA followed by Tukey's test (for A–E) was adopted for statistical analysis. * p < .05, ** p < .01, and *** p < .001.

Article Snippet: The PVDF membranes were then blocked with 5% skim milk for 1 h and treated with primary antibodies against KLF13 (PA5‐80754, 1:1000, Thermo Fisher Scientific), SH2B1 (12226‐1‐AP, 1:500, Proteintech), IRS1 (ab131487, 1:2000, Abcam), HK2 (22029‐1‐AP, 1:1500, Proteintech), GLUT1 (ab150299, 1:200, Abcam), PDK1 (ab202468, 1:2000, Abcam), ENO1 (ab227978, 1:1000, Abcam), LDHA (PA5‐27406, 1:3000, Thermo Fisher Scientific), p‐PI3K (PA5‐104853,1:500, Thermo Fisher Scientific), PI3K (PA5‐29220, 1:500, Thermo Fisher Scientific), p‐AKT (44‐621G, 1:1500, Thermo Fisher Scientific), AKT (MA5‐14916, 1:2000, Thermo Fisher Scientific), CD63 (ab134045, 1:1500, Abcam), TSG101 (ab125011, 1:2000, Abcam), CD9 (ab236630, 1:1500, Abcam), CD81 (ab155760, 1:1000, Abcam), GM130 (PA1‐077, 1:500, Thermo Fisher Scientific), Calnexin (ab22595, 1:1000, Abcam), Annexin V (ab108321, 1:1500, Abcam), Albumin (ab207327, 1:2000, Abcam), JAK2 (ab108596, 1:3000, Abcam), His (ab9108, 1:1000, Abcam), GST (ab111947, 1:1000, Abcam), and β‐actin (ab8227, 1:1000, Abcam) overnight at 4°C.

Techniques: Transfection, Over Expression, Plasmid Preparation, Quantitative Proteomics, Western Blot, Expressing, Immunohistochemical staining, Staining, Quantitative RT-PCR