primary antibodies against glut1  (Bioss)

Journal: Poultry Science

Article Title: Chromium picolinate alleviates heat stress-induced breast muscle glucose and lipid metabolism disorders in broiler chickens

doi: 10.1016/j.psj.2025.105704

Figure Lengend Snippet: Effects of chromium picolinate on breast muscle glycolipid metabolism genes expression of broilers under heat stress. (A) Western blot analysis. (B) GLUT1. (C) PI3K. (D) GS. (E) PPARα. (F) CPT-1. (G) LPL. Abbreviation: GLUT1: glucose transporter 1, PI3K: phosphatidylinositol 3 kinase, GS: glycogen synthase, PPARα: peroxisome proliferators-activated receptors α, CPT-1; carnitine palmitoyl transterase-1, LPL: lipoprotein lipase. In the same rank, a, b, c values with different small letter superscripts mean significant difference ( P < 0.05).

Article Snippet: Primary antibodies against GLUT1 (1:1000, AWA61570 , Abiowell), PI3K (1:1000, AWA63394 , Abiowell), GS (1:1000, AWA43660 , Abiowell), CPT1 (1:1000, AWA63836 , Abiowell), LPL (1:1000, AWA61104 , Abiowell), PPARα (1:500, BS-23398, BIOSS) and β-actin (1:2000, AWA80002 ; Abiowell) were diluted in antibody diluent (AWB0200c; Abiowell) and incubated with the membrane overnight at 4°C.

Techniques: Expressing, Western Blot

Journal: Life Metabolism

Article Title: Hyaluronidase-1 mediates postprandial suppression of hepatic gluconeogenesis

doi: 10.1093/lifemeta/loaf016

Figure Lengend Snippet: Hepatic Hyal1 overexpression reduces metabolites in the gluconeogenesis pathway. (a) Hepatic metabolites from gluconeogenesis pathway in control and LHY TG mice kept on LFD. (b) Hepatic metabolites from gluconeogenesis pathway in control and LHY TG mice kept on 12-week HFD. (c) Hepatic metabolites from the TCA cycle pathway in control and LHY TG mice kept on LFD. (d) Hepatic metabolites from TCA cycle pathway in control and LHY TG mice kept on 12-week HFD. In panels (a) and (c), n = 8 mice for each group. In panels (b) and (d), n = 16 mice for the control group, n = 13 mice for the LHY TG group. Abbreviations of metabolites in panels (a)–(d): Glu: glucose; Fru: fructose; G6P: glucose-6-phosphate; F6P: fructose-6-phosphate; GA3P: glyceraldehyde-3-phosphate; G3P: glycerol-3-phosphate; 3PG/2PG: 3-phosphoglycerate/2-phosphoglycerate; PEP: phosphoenolpyruvate; OAA: oxaloacetic acid; Cit/ICT: citrate/isocitrate; AKG: α-keto glutarate. (e) Western blot analysis of GLUT1 and GLUT2 in liver lysates of control or LHY TG mice on LFD or HFD. Densitometry quantifications are shown on the right. Two-way ANOVA followed by post-hoc Sidak multiple comparison tests were performed. n = 3 and 4 mice for LFD and HFD groups, respectively. (f) Illustrations of metabolites changed in Hyal1 -overexpressing liver from HFD mice in gluconeogenesis pathway. The green arrow indicates a statistically significant reduction in abundance in LHY TG liver lysates in comparison to control liver lysates. ND: not determined. All results are shown as mean ± SEM. ns, not significant. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Protein abundance was detected using primary antibodies against GLUT1 (Novus Biologicals, NB110-39113SS, 1:1,000 dilution), GLUT2 (Novus Biologicals, NBP2-22218SS, 1:500 dilution), PEPCK/PCK2 (Abclonal, A4466, 1:1,000 dilution), PC (Abclonal, A8980, 1:1,000 dilution), O- GlcNAc (Novus Biologicals, NB300-524SS, 1:1,000 dilution), total OXPHOS (Abcam, ab110413, 1:1,000 dilution), ATP5B (Novusbio, NBP3-15354, 1:1,000 dilution), HSP60 (Novusbio, NBP1-77397SS, 1:1,000 dilution), or β-Actin (Cell Signaling Technology, 3700S, 1:1,000 dilution).

Techniques: Over Expression, Control, Western Blot, Comparison

Journal: International Dental Journal

Article Title: Pexidartinib Inhibits Macrophage Senescence Through Glycolysis in Periodontitis Microenvironment

doi: 10.1016/j.identj.2025.100843

Figure Lengend Snippet: Macrophage senescence is accompanied by enhanced glycolytic expression in vivo and in vitro. A, An F4/80 antibody (pink), a p16 antibody (green), and a GLUT antibody (red) were used to stain gingival tissues of the N group and PDs group for immunofluorescence. The nuclei were stained with DAPI (blue). White arrows point to the triple-positive cells.Scale bar, 20 μm and 10 μm. B, Quantification of CSF-1R+ p16+ F4/80+cells. C, Effect of Pg -LPS on the expression of GLUT1,HK2 mRNA in RAW264.7 cell. D, Effect of Pg -LPS on the expression of GLUT1,HK2 protein in RAW264.7 cell. N, normal mice; PDs, periodontitis mice; N, control cells; L/LPS, RAW264.7 cells cultured in Pg -LPS(1μg/ml) for 24h. Data were represented as the mean ± SD (n=3) and analysed relative to the control group, * P < .05, ** P < .01, *** P < .001, **** P < .0001 vs N

Article Snippet: Sections were incubated with primary antibodies against F4/80 (1:1,000, Abcam, UK, Ab300421 ), CSF-1R (1:200, Immunoway, USA, YT0881), p16 (1:200, HUABIO, China, SR34-02), and GLUT1 (1:200, Immunoway, USA, YT1928).

Techniques: Expressing, In Vivo, In Vitro, Staining, Immunofluorescence, Control, Cell Culture

Journal: International Dental Journal

Article Title: Pexidartinib Inhibits Macrophage Senescence Through Glycolysis in Periodontitis Microenvironment

doi: 10.1016/j.identj.2025.100843

Figure Lengend Snippet: Glycolysis modulates Pg -LPS-induced senescence in RAW264.7 cell. A, Impact of 2-DG/GM-CSF pretreatment on HK2,GLUT1 protein expression in RAW264.7 cells stimulated by Pg -LPS. B, Effect of 2-DG/GM-CSF pretreatment on the expression of HK2, and GLUT1 mRNA in RAW264.7 cells stimulated by Pg -LPS. C, The senescent cells were visualised using SA-β-Gal staining. Senescent RAW264.7 cells were symbolised by blue. Scale bar, 100 μm. D, Effect of 2-DG/GM-CSF pretreatment on the expression of SASP factor mRNA in RAW264.7 cells stimulated by Pg -LPS. E, Effect of 2-DG/GM-CSF pretreatment on the p16, p21 protein in RAW264.7 cells stimulated by Pg -LPS. N, control cells; L/LPS, RAW264.7 cultured in Pg -LPS(1μg/ml) for 24h; L+2/LPS+2-DG, 1mM 2-DG pretreatment group; L+G/LPS+GM-CSF, 100ng/ml GM-CSF pretreatment group. Data were represented as the mean ± SD (n=3) and analysed relative to the Pg -LPS group, * P < .05, ** P < .01, *** P < .001, **** P < .0001vs. LPS

Article Snippet: Sections were incubated with primary antibodies against F4/80 (1:1,000, Abcam, UK, Ab300421 ), CSF-1R (1:200, Immunoway, USA, YT0881), p16 (1:200, HUABIO, China, SR34-02), and GLUT1 (1:200, Immunoway, USA, YT1928).

Techniques: Expressing, Staining, Control, Cell Culture

Journal: International Dental Journal

Article Title: Pexidartinib Inhibits Macrophage Senescence Through Glycolysis in Periodontitis Microenvironment

doi: 10.1016/j.identj.2025.100843

Figure Lengend Snippet: PLX3397 Inhibited RAW264.7 cell senescence through glycolysis. A.Impact of PLX3397 pretreatment on the expression of GLUT1,HK2 protein in RAW264.7 cells stimulated by Pg -LPS. B. Effect of PLX3397 pretreatment on the manifestation of GLUT1, and HK2 mRNA in RAW264.7 cells stimulated by Pg -LPS. N, control cells; L/LPS, RAW264.7 cultured in Pg -LPS(1μg/ml) for 24h; L+P/LPS+PLX3397,500nM PLX3397 pretreatment group. Data were represented as the mean ± SD (n=3) and analysed relative to the Pg -LPS group, * P < .05, ** P < .01, *** P < .001, **** P < .0001vs. LPS

Article Snippet: Sections were incubated with primary antibodies against F4/80 (1:1,000, Abcam, UK, Ab300421 ), CSF-1R (1:200, Immunoway, USA, YT0881), p16 (1:200, HUABIO, China, SR34-02), and GLUT1 (1:200, Immunoway, USA, YT1928).

Techniques: Expressing, Control, Cell Culture

Journal: The Journal of Clinical Investigation

Article Title: GLUT3 promotes macrophage signaling and function via RAS-mediated endocytosis in atopic dermatitis and wound healing

doi: 10.1172/JCI170706

Figure Lengend Snippet: ( A and B ) mRNA expression levels of GLUT and SGLT transporter isoforms in BMDMs ( A ) and THP-1 cells ( B ) cells in unstimulated macrophages (green) and after treatment with classic M1 (purple) or alternative M2 (yellow) polarization stimuli for 24 hours. Expression normalized to that of β-actin ( ACTB ) ( n = 3 biological replicates). ( C and D ) Western blot assessing expression of GLUT1 and GLUT3 with the indicated polarization stimuli in BMDMs ( C ) and THP-1 ( D ). iNOS and p-STAT1, M1 polarization markers; Arg1 and MRC1, M2 polarization markers; Hsp90, loading control. ( E ) GLUT1 ( Slc2a1 ) and GLUT3 ( Slc2a3 ) expression in primary human CD14 + peripheral blood monocyte–derived macrophages after treatment with indicated polarization stimuli. ( F ) mRNA expression levels of M1 ( Nos2 , Tnfa , and Il1b ) and M2 ( Arg1 , Retnla , and Chil3l3 ) markers in WT ( n = 12) and LysM-Cre Glut1 fl/fl (GLUT1 KO) ( n = 12) BMDMs after the indicated polarization stimuli ( n = 4 biological replicates). ( G ) mRNA expression levels of M1 and M2 markers in WT ( n = 12) and LysM-Cre Glut3 fl/fl (GLUT3 KO) ( n = 12) BMDMs after the indicated polarization stimuli ( n = 4 biological replicates). ( H ) 2-Deoxy-D-glucose uptake in WT, LysM-Cre Glut1 fl/fl , and LysM-Cre Glut3 fl/fl BMDMs after the indicated polarization stimuli. Fold change represents uptake relative to uptake in unstimulated BMDMs from the same mouse. Data shown as mean ± SEM ( n = 2 biological replicates). ( I ) Pyruvate and ATP levels in WT, LysM-Cre Glut1 fl/fl , and LysM-Cre Glut3 fl/fl BMDMs after the indicated polarization stimuli. P values were calculated by 2-way ANOVA with Dunnett’s test ( A and B ), 1-way ANOVA with Dunnett’s test ( E and I ), or 2-tailed t test ( H ). * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001.

Article Snippet: Cells were then incubated with primary antibodies against GLUT1 (MilliporeSigma, 07-1401), GLUT3 (Santa Cruz Biotechnology, sc-30107), and EEA1 (Cell Signaling Technology, 48453) in blocking buffer overnight at 4°C, followed by 1 hour of incubation with fluorescent dye–labeled secondary antibodies (Thermo Fisher Scientific, A11005 or A11008).

Techniques: Expressing, Western Blot, Derivative Assay

Journal: The Journal of Clinical Investigation

Article Title: GLUT3 promotes macrophage signaling and function via RAS-mediated endocytosis in atopic dermatitis and wound healing

doi: 10.1172/JCI170706

Figure Lengend Snippet: ( A ) Violin plots showing relative expression of GLUT1 and GLUT3 in alternative macrophages (Mac2) from single-cell RNA-seq profiles of CD45 + cells from patients with psoriasis vulgaris ( n = 8), atopic dermatitis ( n = 7), or healthy controls ( n = 7). ( B ) Representative photos after calcipotriol administration in WT, LysM-Cre Glut3 fl/fl (GLUT3 KO), and LysM-Cre Glut1 fl/fl (GLUT1 KO) mice on day 8. ( C ) Hematoxylin and eosin–stained sections of mouse skin treated with calcipotriol analyzed on day 13 in WT, LysM-Cre Glut3 fl/fl , and LysM-Cre Glut1 fl/fl mice. Scale bars: 100 μm. ( D ) Thickness of calcipotriol-treated ear and back in WT ( n = 11 for ear and n = 6 for back), LysM-Cre Glut3 fl/fl ( n = 12 for ear and n = 8 for back), and LysM-Cre Glut1 fl/fl ( n = 10 for ear and n = 4 for back) mice. ( E and F ) mRNA expression levels in calcipotriol-treated ear in WT ( n = 3), LysM-Cre Glut3 fl/fl ( n = 4), and LysM-Cre Glut1 fl/fl ( n = 3) mice. Pan-macrophage marker ( Adgre1 [F4/80]), M1 markers ( Nos2 and Tnfa ) ( E ), and M2 markers ( Arg1 , Mrc1 , and Retnla ) ( F ) were observed. ( G ) Representative immunofluorescent staining of Arg1 (green) and F4/80 (red) in the back skin of calcipotriol treated mice (day 8). Scale bars: 50 μm. (Right) Quantification of M2 macrophages at wound sites (day 8). The number of F4/80 + Arg1 + (M2) cells present relative to the total number of F4/80 + cells. Data shown as mean ± SEM. P values were calculated by 1-way ANOVA with Dunnett’s test. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001.

Article Snippet: Cells were then incubated with primary antibodies against GLUT1 (MilliporeSigma, 07-1401), GLUT3 (Santa Cruz Biotechnology, sc-30107), and EEA1 (Cell Signaling Technology, 48453) in blocking buffer overnight at 4°C, followed by 1 hour of incubation with fluorescent dye–labeled secondary antibodies (Thermo Fisher Scientific, A11005 or A11008).

Techniques: Expressing, RNA Sequencing Assay, Staining, Marker

Journal: The Journal of Clinical Investigation

Article Title: GLUT3 promotes macrophage signaling and function via RAS-mediated endocytosis in atopic dermatitis and wound healing

doi: 10.1172/JCI170706

Figure Lengend Snippet: ( A ) Violin plots showing relative expression of GLUT1 and GLUT3 in alternative macrophages (Mac2) from single-cell RNA-seq profiles of indicated tissues from diabetic patients with no foot ulcer ( n = 6), healing diabetic foot ulcer ( n = 7), healthy nondiabetic skin ( n = 10), or nonhealing diabetic foot ulcer ( n = 4). Transcriptomic data are from Theocharidis et al. . DFU, diabetic foot ulcer. ( B ) Representative immunofluorescent staining of a patient biopsy specimen of a wound bed for CD68 (red) and GLUT3 (green). Arrows indicate cells expressing both CD68 and GLUT3 in the wound bed (see ). Scale bar: 20 μm. ( C ) Representative photos of wound site in WT, LysM-Cre Glut3 fl/fl (GLUT3 KO), and LysM-Cre Glut1 fl/fl (GLUT1 KO) mice 6 days after injury. ( D ) Measurements of wound diameter on day 6 in WT ( n = 12), LysM-Cre Glut3 fl/fl ( n = 12), and LysM-Cre Glut1 fl/fl ( n = 7) mice (see ). ( E ) Quantification of M2 macrophages at wound sites (day 6). The number of F4/80 + Arg1 + (M2) cells present relative to the total number of F4/80 + cells (see ). ( F ) Representative immunofluorescent staining for Arg1 (green), F4/80 (red), and with DAPI (blue) in the wound site (day 6). Scale bars: 50 μm. ( G and H ) mRNA expression levels of M0/M1 markers ( Adgre1 [F4/80], Nos2 , and Tnfa ) ( E ) or M2 markers ( Arg1 , Mrc1 , and Retnla ) ( F ) in WT ( n = 3), LysM-Cre Glut3 fl/fl ( n = 3), and LysM-Cre Glut1 fl/fl ( n = 3) mice. Data shown as mean ± SEM. P values were calculated by 1-way ANOVA with Dunnett’s test. * P ≤ 0.05; *** P ≤ 0.001; **** P ≤ 0.0001.

Article Snippet: Cells were then incubated with primary antibodies against GLUT1 (MilliporeSigma, 07-1401), GLUT3 (Santa Cruz Biotechnology, sc-30107), and EEA1 (Cell Signaling Technology, 48453) in blocking buffer overnight at 4°C, followed by 1 hour of incubation with fluorescent dye–labeled secondary antibodies (Thermo Fisher Scientific, A11005 or A11008).

Techniques: Expressing, RNA Sequencing Assay, Staining

Journal: The Journal of Clinical Investigation

Article Title: GLUT3 promotes macrophage signaling and function via RAS-mediated endocytosis in atopic dermatitis and wound healing

doi: 10.1172/JCI170706

Figure Lengend Snippet: ( A ) Representative immunofluorescence image of THP-1 cells labeled for GLUT1 (upper panel, green), GLUT3 (lower panel, green), EEA1 (red), and with DAPI (blue). Scale bars: 10 μm. ( B – D ) Western blot analysis of the expression of GLUT3, GLUT1, p-STAT6, and STAT6 in the isolated plasma membrane (PM) and endosome fraction from WT and LysM-Cre Glut3 fl/fl (GLUT3 KO) BMDMs ( B ), THP-1 cells ( C ), and Raw 264.7 cells ( D ). Na + /K + -ATPase and EEA1 are fractionation controls for the plasma membrane and endosome, respectively. ( E ) Western blot analysis of the expression of p-STAT6 and t-STAT6 in BMDMs with and without IL-4 (30 minutes) and with and without Dynasore.

Article Snippet: Cells were then incubated with primary antibodies against GLUT1 (MilliporeSigma, 07-1401), GLUT3 (Santa Cruz Biotechnology, sc-30107), and EEA1 (Cell Signaling Technology, 48453) in blocking buffer overnight at 4°C, followed by 1 hour of incubation with fluorescent dye–labeled secondary antibodies (Thermo Fisher Scientific, A11005 or A11008).

Techniques: Immunofluorescence, Labeling, Western Blot, Expressing, Isolation, Membrane, Fractionation

Journal: The Journal of Clinical Investigation

Article Title: GLUT3 promotes macrophage signaling and function via RAS-mediated endocytosis in atopic dermatitis and wound healing

doi: 10.1172/JCI170706

Figure Lengend Snippet: ( A and B ) Western blot (WB) of IL-4Rα and γ c chain in the plasma membrane (PM) and endosomal fractions from WT, LysM-Cre Glut3 fl/fl (GLUT3 KO) BMDMs ( A ), and THP-1 cells transduced with control or GLUT3 shRNA ( B ). Na + /K + -ATPase and EEA1, fractionation controls. Mean of IL-4Rα and γ c chain levels relative to EEA1 from quantification of WB ( n = 3 biological replicates; , C and F). ( C ) HEK293T cells were transfected with the indicated GLUT allele, and GLUT1 (Thermo Fisher Scientific, MA1-37783) or GLUT3 (Abcam, ab15311) was immunoprecipitated. RAS was detected by Western blotting. Normal mouse/rabbit IgG, IP controls. ( D ) HEK293T cells were transfected with the indicated GLUT allele and immunoprecipitation performed after serum starvation (serum-free, SF) or refeeding as indicated. ( E ) HEK293T cells were transfected with the indicated GLUT3 allele and GLUT3 alleles were FLAG immunoprecipitated; RAS was detected by Western blotting. IgG indicates a normal mouse IgG control. ( F ) The indicated GST fusion protein was bound to glutathione-agarose and incubated with GDP- or GTP-bound (GMP-PNP) KRAS as indicated. Bound proteins were eluted and assessed by Western blotting. The GST blot was stripped and probed for RAS to ensure even loading. ( G ) Levels of p-STAT6 after expression of indicated shRNA-resistant GLUT3 allele and shRNA of endogenous GLUT3 in THP-1 cells. ( H ) THP-1 were transduced with the indicated shRNA and shRNA-resistant GLUT3 allele and then the indicated M2 marker ( MRC1 , TGM2 ) was assessed by qRT-PCR after IL-4 stimulation (24 hours). P values were calculated by 1-way ANOVA with Dunnett’s test. *** P ≤ 0.001, **** P ≤ 0.0001.

Article Snippet: Cells were then incubated with primary antibodies against GLUT1 (MilliporeSigma, 07-1401), GLUT3 (Santa Cruz Biotechnology, sc-30107), and EEA1 (Cell Signaling Technology, 48453) in blocking buffer overnight at 4°C, followed by 1 hour of incubation with fluorescent dye–labeled secondary antibodies (Thermo Fisher Scientific, A11005 or A11008).

Techniques: Western Blot, Membrane, Transduction, shRNA, Fractionation, Transfection, Immunoprecipitation, Incubation, Expressing, Marker, Quantitative RT-PCR