mouse monoclonal glut3 abs sc74399  (Santa Cruz Biotechnology)

Journal: Cells

Article Title: TGFβ1 Restores Energy Homeostasis of Human Trophoblast Cells Under Hyperglycemia In Vitro by Inducing PPARγ Expression, AMPK Activation, and HIF1α Degradation

doi: 10.3390/cells14010045

Figure Lengend Snippet: TGFβ1 differentially regulates glucose transporters expression, lactate, and ATP production: ( A–E ) JEG-3 cells were cultured for 24 h in media containing normal-glucose (5 mM) or high-glucose (25 mM) concentrations in the absence or presence of TGFβ1 at 50 ng/mL. ( A ) Representative images of GLUT1, GLUT3, and b-actin protein detection, as assessed by western blot. Graphical analysis showing the relative expression of GLUT1 ( B ) and GLUT3 ( C ) at 5 mM and 25 mM glucose. ( D ) Quantitation of lactate production was assessed using the Lactate-Glo TM assay kit. ( E ) Quantitation of ATP production was assessed using the ATP luminescence detection assay kit. Data are expressed as nM of ATP and lactate ( n = 3). ( F ) Quantitation of relative cell viability was assessed using the MTT assay ( n = 3). Each bar represents the mean ± SD from at least two independent experiments. * p < 0.05 indicates a significant difference between the cell groups, and ns = nonsignificant difference.

Article Snippet: The Total OXPHOS Rodent WB Antibody Cocktail (ab110413) was sourced from Abcam (Cambridge, UK), and the mouse monoclonal GLUT3 Abs (Sc-74399) from Santa Cruz Biotechnology (Cambridge, MA, USA).

Techniques: Expressing, Cell Culture, Western Blot, Quantitation Assay, Detection Assay, MTT Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Berberine Modulates Macrophage Activation by Inducing Glycolysis.

doi: 10.4049/jimmunol.2100716

Figure Lengend Snippet: FIGURE 2. Effect of berberine and LPS on glycolytic enzymes in BMDMs. (A and B) BMDMs treated with 3.3 mM or 1 mg/ml LPS for 6 and 24 h in DMEM (25 mM glucose) were used to detect protein levels involved in glycolysis by Western blot. A part of the protein was taken out to blot Glut1 without boiling. (C) Relative RNA levels of Glut1, Glut3, and Glut4 in BMDMs measured by qPCR. (D) RNA level ratio of BMDMs treated with berberine (6.6 mM) for 24 h to cells treated with DMSO. (E) RNA level ratio of BMDMs treated with 200 ng/ml LPS to cells treated with saline for 6 h. (F and G) HK activities of BMDMs treated with 6.6 mM berberine or 200 ng/ml LPS were measured in vitro. (H) HeLa cells were transfected with FLAG-tagged HK1 or FLAG-HK2 plasmids. Twenty-four hours after transfection, cells were treated with berberine and the subcellular localization of exogenous HK1/2 was stained by FLAG Ab. Images are shown at original magnification ×400. *p < 0.05, **p < 0.01, ***p < 0.001. BBR, berberine.

Article Snippet: Abs for Glut3 (sc-74399), Glut4 (sc53566), and HXK1 (sc-46695) were from Santa Cruz Biotechnology.

Techniques: Western Blot, Saline, In Vitro, Transfection, Staining