Journal: Cell Reports Methods

Article Title: In situ microwave fixation provides an instantaneous snapshot of the brain metabolome

doi: 10.1016/j.crmeth.2023.100455

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Article Snippet: GLUT3, Rabbit , Alomone , Cat# AGT-023; RRID:AB_2756644.

Techniques:

Journal: Journal of Translational Medicine

Article Title: Hypoxic BMSCs accelerate femur fracture healing via the METTL3/SLC2A3 m6A-glycolysis axis

doi: 10.1186/s12967-025-07510-2

Figure Lengend Snippet: SLC2A3 enhances osteogenic capacity through the glycolytic pathway. ( A ) Immunofluorescence assay showed the expression level of SLC2A3. ( B - C ) WB analysis of SLC2A3, Runx2, OCN, and Osx expression levels in groups. ( D ) ELISA analysis of ALP levels in groups. ( E ) Alizarin Red staining illustrating osteogenic capacity. ( F ) ATP levels. ( G ) Glucose uptake. ( H ) WB analysis of HK2, PKM2, and LDHA expression levels in groups. ( I ) The detection of extracellular acidification rate (ECAR). ** P < 0.01, *** P < 0.001, one-way ANOVA

Article Snippet: Membranes were incubated with primary antibodies against SLC2A3 (#A03259-2, 1:1000, BOSTER, Wuhan, China), runt-related transcription factor-2 (Runx2, #ab236639, 1:1000, Abcam, Cambridge, MA, USA), osteocalcin (OCN, #ab93876, 1:1000), Osterix (Osx, #A02077-1, 1:1000), hexokinase 2 (HK2, #ab209847, 1:2000), pyruvate kinase muscle isozyme 2 (PKM2, #ab150377, 1:2000), lactate dehydrogenase A (LDHA, #ab112996, 1:1000), METTL3 (#ab195352, 1:1000), METTL4 (#ab261735, 1:1000), METTL4 (#ab309096, 1:1000), and ACTB (#ab8226, 1:5000).

Techniques: Immunofluorescence, Expressing, Enzyme-linked Immunosorbent Assay, Staining

Journal: Journal of Translational Medicine

Article Title: Hypoxic BMSCs accelerate femur fracture healing via the METTL3/SLC2A3 m6A-glycolysis axis

doi: 10.1186/s12967-025-07510-2

Figure Lengend Snippet: METTL3 promotes osteogenic capacity through glycolysis. ( A ) qPCR analysis of the expression level of METTL3 in Ctrl and Hypoxia groups. ( B , D ) WB analysis of the expression levels of METTL3, METTL4, METTL14, SLC2A3, Runx2, OCN, and Osx in groups. ( C , E ) Immunofluorescence assay showed the expression level of METTL3 and SLC2A3. (F) WB analysis of Runx2, OCN, and Osx expression levels. ( G ) ELISA analysis of ALP levels in groups. ( H ) Glucose uptake. ( I ) ATP level. ( J ) Alizarin Red staining detects osteogenic capacity. (K) WB analysis of HK2, PKM2, and LDHA expression levels in groups. ( L ) The detection of ECAR. * P < 0.05, ** P < 0.01, *** P < 0.001, Student’s t -test

Article Snippet: Membranes were incubated with primary antibodies against SLC2A3 (#A03259-2, 1:1000, BOSTER, Wuhan, China), runt-related transcription factor-2 (Runx2, #ab236639, 1:1000, Abcam, Cambridge, MA, USA), osteocalcin (OCN, #ab93876, 1:1000), Osterix (Osx, #A02077-1, 1:1000), hexokinase 2 (HK2, #ab209847, 1:2000), pyruvate kinase muscle isozyme 2 (PKM2, #ab150377, 1:2000), lactate dehydrogenase A (LDHA, #ab112996, 1:1000), METTL3 (#ab195352, 1:1000), METTL4 (#ab261735, 1:1000), METTL4 (#ab309096, 1:1000), and ACTB (#ab8226, 1:5000).

Techniques: Expressing, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Staining

Journal: Journal of Translational Medicine

Article Title: Hypoxic BMSCs accelerate femur fracture healing via the METTL3/SLC2A3 m6A-glycolysis axis

doi: 10.1186/s12967-025-07510-2

Figure Lengend Snippet: BMSC hypoxia promotes METTL3, which facilitates osteogenic capacity by activating the m 6 A modification of SLC2A3. ( A ) After treating cells with 1 µM SAH, RIP-qPCR was employed to detect the m6A modification level on SLC2A3 mRNA. ( B - D ) QPCR, WB and immunofluorescence assay showed the expression level of SLC2A3. ( E ) The detection of ECAR. ( F , J ) WB analysis of METTL3, SLC2A3, Runx2, OCN, and Osx expression levels in transfections. ( G ) ELISA analysis of ALP levels in groups. ( H ) Glucose uptake. ( I ) ATP level. ( K ) Alizarin Red staining detects osteogenic capacity. ( L ) WB analysis of HK2, PKM2, and LDHA expression levels in transfections. ( M ) The detection of ECAR. ***P < 0.001, one-way ANOVA

Article Snippet: Membranes were incubated with primary antibodies against SLC2A3 (#A03259-2, 1:1000, BOSTER, Wuhan, China), runt-related transcription factor-2 (Runx2, #ab236639, 1:1000, Abcam, Cambridge, MA, USA), osteocalcin (OCN, #ab93876, 1:1000), Osterix (Osx, #A02077-1, 1:1000), hexokinase 2 (HK2, #ab209847, 1:2000), pyruvate kinase muscle isozyme 2 (PKM2, #ab150377, 1:2000), lactate dehydrogenase A (LDHA, #ab112996, 1:1000), METTL3 (#ab195352, 1:1000), METTL4 (#ab261735, 1:1000), METTL4 (#ab309096, 1:1000), and ACTB (#ab8226, 1:5000).

Techniques: Modification, Immunofluorescence, Expressing, Transfection, Enzyme-linked Immunosorbent Assay, Staining

Journal: Journal of Translational Medicine

Article Title: Hypoxic BMSCs accelerate femur fracture healing via the METTL3/SLC2A3 m6A-glycolysis axis

doi: 10.1186/s12967-025-07510-2

Figure Lengend Snippet: BMSC hypoxia promotes METTL3 and facilitates osteogenic capacity by activating SLC2A3 m 6 A modification through the glycolytic pathway. ( A ) RIP analysis of the binding relationship between SLC2A3 and METTL3. ( B ) Pull-down analysis of the association between SLC2A3 and METTL3. ( C - D ) WB analysis of SLC2A3, Runx2, OCN, and Osx expression levels in transfections. ( E ) Glucose uptake. ( F ) ATP levels. ( G ) WB analysis of HK2, PKM2, and LDHA expression levels in transfections. ( H ) Alizarin Red staining to detect osteogenic capacity. *** P < 0.001, Student’s t-test

Article Snippet: Membranes were incubated with primary antibodies against SLC2A3 (#A03259-2, 1:1000, BOSTER, Wuhan, China), runt-related transcription factor-2 (Runx2, #ab236639, 1:1000, Abcam, Cambridge, MA, USA), osteocalcin (OCN, #ab93876, 1:1000), Osterix (Osx, #A02077-1, 1:1000), hexokinase 2 (HK2, #ab209847, 1:2000), pyruvate kinase muscle isozyme 2 (PKM2, #ab150377, 1:2000), lactate dehydrogenase A (LDHA, #ab112996, 1:1000), METTL3 (#ab195352, 1:1000), METTL4 (#ab261735, 1:1000), METTL4 (#ab309096, 1:1000), and ACTB (#ab8226, 1:5000).

Techniques: Modification, Binding Assay, Expressing, Transfection, Staining

Journal: Journal of Translational Medicine

Article Title: Hypoxic BMSCs accelerate femur fracture healing via the METTL3/SLC2A3 m6A-glycolysis axis

doi: 10.1186/s12967-025-07510-2

Figure Lengend Snippet: SLC2A3 overexpression promotes fracture healing. ( A ) X-ray images of bone defects in rat models of the different groups after surgery ( n = 3). ( B ) Gross observation of the surgical site in Sprague–Dawley rats ( n = 3). ( C ) Micro-CT analysis of fracture tissues ( n = 3). ( D ) WB analysis of SLC2A3, METTL3, Runx2, HK2, PKM2, and LDHA protein expression levels ( n = 3)

Article Snippet: Membranes were incubated with primary antibodies against SLC2A3 (#A03259-2, 1:1000, BOSTER, Wuhan, China), runt-related transcription factor-2 (Runx2, #ab236639, 1:1000, Abcam, Cambridge, MA, USA), osteocalcin (OCN, #ab93876, 1:1000), Osterix (Osx, #A02077-1, 1:1000), hexokinase 2 (HK2, #ab209847, 1:2000), pyruvate kinase muscle isozyme 2 (PKM2, #ab150377, 1:2000), lactate dehydrogenase A (LDHA, #ab112996, 1:1000), METTL3 (#ab195352, 1:1000), METTL4 (#ab261735, 1:1000), METTL4 (#ab309096, 1:1000), and ACTB (#ab8226, 1:5000).

Techniques: Over Expression, Micro-CT, Expressing

Journal: The Journal of Neuroscience

Article Title: Energy Requirements of Odor Transduction in the Chemosensory Cilia of Olfactory Sensory Neurons Rely on Oxidative Phosphorylation and Glycolytic Processing of Extracellular Glucose

doi: 10.1523/JNEUROSCI.2640-16.2017

Figure Lengend Snippet: Olfactory neuron cilia and sustentacular cell microvilli express the high affinity GLUT3 glucose transporter. A, Immunocytochemistry for GLUT3 without (left) or with (right) preincubation of the primary antibody (Alpha Diagnostics anti-GLUT3 antibody) with its antigenic peptide in rat OSNs, as a control for the specificity of the antibody. The left three panels show a neuron labeled for OMP, GLUT3, and a merge of both; the two right panels show the control. B, Immunocytochemistry (Sigma anti-GLUT3 antibody) for GLUT3 of an isolated rat OSN. C, Isolated toad OSN (Sigma anti-GLUT3 antibody). D, Rat SCs (Sigma anti-GLUT3 antibody). GLUT3-expressing cilia are indicated by arrowheads. C, D, Insets (3×) correspond to the areas denoted in the figures. Nuclei are labeled blue (TOPRO). Scale bars, 10 μm.

Article Snippet: For double labeling, immunofluorescence cells were incubted overnight with the primary rabbit anti-GLUT3 and goat anti-OMP (1:200, Catalog #544–10001 Wako Chemicals), washed and then incubated 2 h at room temperature with the secondary donkey anti-rabbit Alexa-488 (1:500, A-21206, Invitrogen) and donkey anti-goat Alexa-594 (1:500, ab150136, Abcam).

Techniques: Immunocytochemistry, Labeling, Isolation, Expressing

Journal: Journal of animal science and biotechnology

Article Title: Dietary supplementation with pterostilbene activates the PI3K-AKT-mTOR signalling pathway to alleviate progressive oxidative stress and promote placental nutrient transport.

doi: 10.1186/s40104-024-01090-9

Figure Lengend Snippet: Fig. 5 Effect of PTE on placental nutrient transport capacity. A–C Fatty acid transporter FATP1 and glucose transporter GLUT3, 200× analysis of IF staining. Data are expressed as mean ± SD (n = 3 for each group). *P < 0.05, compared to the control group

Article Snippet: Anti-GLUT3 (Wanlei WL05064) antibody was purchased from Wanleibio (Shenyang, China).

Techniques: Staining, Control