Journal: Cellular and Molecular Neurobiology

Article Title: Sevoflurane Induces Learning and Memory Impairment in Young Mice Through a Reduction in Neuronal Glucose Transporter 3

doi: 10.1007/s10571-019-00779-0

Figure Lengend Snippet: Sevoflurane decreases GLUT3 protein expression in the hippocampus and temporal lobe. a WB analysis of GLUT1, GLUT4, and GLUT3 protein expression in the hippocampus and temporal lobe of young mice 24 h after the last exposure to sevoflurane. n = 4 for each group. b, c, d Histograms showing the quantification of GLUT1, GLUT4, and GLUT3 blots in the hippocampus and temporal lobe. In the hippocampus, * P < 0.05 versus the Con group. In the temporal lobe, # P < 0.05 versus the Con group. e Fluorescent images showing GLUT3 expression in neurons of the hippocampal CA1 area and temporal lobe (left panel). Quantification of GLUT3 intensity in the CA1 region of the hippocampus and temporal lobe 24 h after the last exposure to sevoflurane (right panel). In the CA1 region of the hippocampus, * P < 0.05 versus the Con group. In the temporal lobe, # P < 0.05 versus the Con group. n = 4 for each group. The data are presented as the mean ± s.e.m

Article Snippet: After incubation with WB-specific blocking solution (5% skimmed milk powder (AS1033, Aspen, China) diluted in TBST), the PVDF membranes were separately incubated with anti-GLUT1 (1:1000, ab652, Abcam), anti-GLUT4 (1:2000, ab654, Abcam), anti-(extracellular) GLUT3 (1:1000, AGT-023, Alomone), anti-Bax (1:800, sc-7480, Santa Cruz), anti-Bcl-2 (1:800, sc-7382, Santa Cruz), anti-caspase-3 (cleaved) (1:100, AB3623, Millipore), anti-poly (ADP-ribose) polymerase (PARP) (cleaved) (1:1000, 5625, Cell Signaling Technology), and anti-β-actin (1:10,000, Aspen) antibodies at 4 °C overnight.

Techniques: Expressing

Journal: Cellular and Molecular Neurobiology

Article Title: Sevoflurane Induces Learning and Memory Impairment in Young Mice Through a Reduction in Neuronal Glucose Transporter 3

doi: 10.1007/s10571-019-00779-0

Figure Lengend Snippet: Sevoflurane reduces GLUT3 protein expression in primary hippocampal neurons. a Primary hippocampal neurons were transfected with LV5-GLUT3. The transfection efficiency was examined by GFP expression (green) 36 h post infection, which was analysed by phase contrast (left panel) and fluorescence microscopy (right panel). b WB analysis of GLUT3 protein expression in primary hippocampal neurons 24 h after the last exposure to sevoflurane. Neurons were normally cultured (the Con group), only transfected with LV5-GLUT3 (the LV5-GLUT3 group), transfected with LV5-GLUT3 and then exposed to sevoflurane (the LV5-GLUT3 + Sev group), and only exposed to sevoflurane (the Sev group). c Immunofluorescence of GLUT3 expression in the somata and dendrites of primary hippocampal neurons 24 h after the last exposure to sevoflurane. The four groups: the LV5 group (neurons transfected with LV5 empty vectors), the LV5 + Sev group (neurons transfected with LV5 empty vectors and then exposed to sevoflurane), the LV5-GLUT3 and the LV5-GLUT3 + Sev group. d Histograms showing the results of GLUT3 blots in neurons among the four groups. n = 5 for each group. * P < 0.05 versus the Con group. # P < 0.05 versus the LV5-GLUT3 group. e Histograms showing the intensity of GLUT3 in the four treatment groups. n = 5 for each group. * P < 0.05 versus the LV5 group. # P < 0.05 versus the LV5-GLUT3 group. The data are presented as the mean ± s.e.m

Article Snippet: After incubation with WB-specific blocking solution (5% skimmed milk powder (AS1033, Aspen, China) diluted in TBST), the PVDF membranes were separately incubated with anti-GLUT1 (1:1000, ab652, Abcam), anti-GLUT4 (1:2000, ab654, Abcam), anti-(extracellular) GLUT3 (1:1000, AGT-023, Alomone), anti-Bax (1:800, sc-7480, Santa Cruz), anti-Bcl-2 (1:800, sc-7382, Santa Cruz), anti-caspase-3 (cleaved) (1:100, AB3623, Millipore), anti-poly (ADP-ribose) polymerase (PARP) (cleaved) (1:1000, 5625, Cell Signaling Technology), and anti-β-actin (1:10,000, Aspen) antibodies at 4 °C overnight.

Techniques: Expressing, Transfection, Infection, Fluorescence, Microscopy, Cell Culture, Immunofluorescence

Journal: Cellular and Molecular Neurobiology

Article Title: Sevoflurane Induces Learning and Memory Impairment in Young Mice Through a Reduction in Neuronal Glucose Transporter 3

doi: 10.1007/s10571-019-00779-0

Figure Lengend Snippet: Sevoflurane inhibits GLUT3 surface expression and GLUT3 mRNA expression in neurons. a PC12 cells were transfected with LV5-GLUT3. The transfection efficiency was examined by GFP expression (green) 36 h post infection, which was analysed by phase contrast (left panel) and fluorescence microscopy (right panel). b The expression levels of GLUT3 protein in PC12 cells infected with LV5-GLUT3 were determined by WB analysis. PC12 cells were normally cultured (the Con group), transfected with LV5 empty vectors (the LV5 group), or transfected with LV5-GLUT3 (the LV5-GLUT3 group). c Flow cytometry analysis of GLUT3 surface expression in PC12 cells 24 h after the last exposure to sevoflurane. Four groups: the Con group, the Sev group (PC12 cells only exposed to sevoflurane), the LV5-GLUT3 group and the LV5-GLUT3 + Sev group (PC12 cells transfected with LV5-GLUT3 and then exposed to sevoflurane). n = 5 for each group. * P < 0.05 versus the Con group. # P < 0.05 versus the LV5-GLUT3 group. d Representative G6P concentrations showing glucose uptake in PC12 cells 24 h after the last exposure to sevoflurane. n = 5 for each group. * P < 0.05 versus the Con group. # P < 0.05 versus the LV5-GLUT3 group. e Representative G6P concentration showing glucose uptake in primary hippocampal neurons 24 h after the last exposure to sevoflurane. n = 5 for each group. * P < 0.05 versus the Con group. # P < 0.05 versus the LV5-GLUT3 group. f Representative quantification of GLUT3 mRNA expression in the hippocampus and temporal lobe 24 h after the last exposure to sevoflurane. n = 4 for each group. In the hippocampus, * P < 0.05 versus the Con group. In the temporal lobe, # P < 0.05 versus the Con group. g Representative quantification of GLUT3 mRNA expression in primary hippocampal neurons 24 h after the last exposure to sevoflurane. n = 5 for each group. * P < 0.05 versus the Con group. # P < 0.05 versus the LV5-GLUT3 group. The data are presented as the mean ± s.e.m

Article Snippet: After incubation with WB-specific blocking solution (5% skimmed milk powder (AS1033, Aspen, China) diluted in TBST), the PVDF membranes were separately incubated with anti-GLUT1 (1:1000, ab652, Abcam), anti-GLUT4 (1:2000, ab654, Abcam), anti-(extracellular) GLUT3 (1:1000, AGT-023, Alomone), anti-Bax (1:800, sc-7480, Santa Cruz), anti-Bcl-2 (1:800, sc-7382, Santa Cruz), anti-caspase-3 (cleaved) (1:100, AB3623, Millipore), anti-poly (ADP-ribose) polymerase (PARP) (cleaved) (1:1000, 5625, Cell Signaling Technology), and anti-β-actin (1:10,000, Aspen) antibodies at 4 °C overnight.

Techniques: Expressing, Transfection, Infection, Fluorescence, Microscopy, Cell Culture, Flow Cytometry, Concentration Assay

Journal: Cellular and Molecular Neurobiology

Article Title: Sevoflurane Induces Learning and Memory Impairment in Young Mice Through a Reduction in Neuronal Glucose Transporter 3

doi: 10.1007/s10571-019-00779-0

Figure Lengend Snippet: Sevoflurane induces neural apoptosis in the hippocampus and temporal lobe. a Representative immunoblots showing the protein expression of Bax, Bcl-2, cleaved caspase-3 and cleaved PARP in the hippocampus and temporal lobe of young mice 24 h after the last exposure to sevoflurane. n = 4 for each group. b, c, d, e Histograms showing the results of Bax, Bcl-2, cleaved caspase-3, and cleaved PARP blots in the hippocampus and temporal lobe. * P < 0.05 versus the Con group. In the temporal lobe, # P < 0.05 versus the Con group. f Representative images showing the flow cytometry analysis of the apoptosis ratio in PC12 cells 24 h after the last exposure to sevoflurane. Early apoptotic populations (PE + 7-AAD − cells) are in the lower-right quadrant; late apoptotic cells (PE + 7-AAD + ) are in the upper-right quadrant in each dot plot. g Representative quantification of the flow cytometry analysis of the apoptosis ratio of PC12 cells 24 h after the last exposure to sevoflurane. h Representative CCK-8 assay showing cell viability in primary hippocampal neurons 24 h after the last exposure to sevoflurane. n = 5 for each group. * P < 0.05 versus the Con group. # P < 0.05 versus the LV5-GLUT3 group. The data are presented as the mean ± s.e.m

Article Snippet: After incubation with WB-specific blocking solution (5% skimmed milk powder (AS1033, Aspen, China) diluted in TBST), the PVDF membranes were separately incubated with anti-GLUT1 (1:1000, ab652, Abcam), anti-GLUT4 (1:2000, ab654, Abcam), anti-(extracellular) GLUT3 (1:1000, AGT-023, Alomone), anti-Bax (1:800, sc-7480, Santa Cruz), anti-Bcl-2 (1:800, sc-7382, Santa Cruz), anti-caspase-3 (cleaved) (1:100, AB3623, Millipore), anti-poly (ADP-ribose) polymerase (PARP) (cleaved) (1:1000, 5625, Cell Signaling Technology), and anti-β-actin (1:10,000, Aspen) antibodies at 4 °C overnight.

Techniques: Western Blot, Expressing, Flow Cytometry, CCK-8 Assay

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