Journal: Brain Structure & Function

Article Title: Somatostatin receptors (SSTR1-5) on inhibitory interneurons in the barrel cortex

doi: 10.1007/s00429-019-02011-7

Figure Lengend Snippet: Localisation of SSTRs in the cortical layers of mice primary somatosensory cortex. SSTR1 showed the most intense signal in L4 and a weaker signal in L2/3 and L5/6. SSTR2 was localised mainly in deep cortical layers (L5-6). SSTR3, SSTR4, and SSTR5 were distributed evenly throughout the cortical layers (column A and C). Column a present representative coronal brain sections through somatosensory cortex immunofluorescently labelled with antibodies specific for SSTR1-SSTR5. Column b presents the validation of antibody specificity. Coronal brain sections were treated with primary antibodies (anti-SSTR1-5) preincubated with control blocking peptides, followed by specific secondary antibodies. The addition of control peptides resulted in the disappearance of the antibody-specific signal. Scale bar 100 μm

Article Snippet: The primary antibodies used in this study were as follows: rabbit anti-SSTR1 polyclonal antibody (1:400, catalogue no. ASR-001, Alomone Labs), rabbit anti-SSTR2 monoclonal antibody (1:2000, catalogue no. ab134152, Abcam), rabbit anti-SSTR3 polyclonal antibody (1:300, catalogue no. ASR-003, Alomone Labs), rabbit anti-SSTR4 polyclonal antibody (1:1000, catalogue no. ab28578, Abcam), and rabbit anti-SSTR5 polyclonal antibody (1:1000, catalogue no. ASR-005, Alomone Labs).

Techniques: Biomarker Discovery, Control, Blocking Assay, Immunofluorescence

Journal: Brain Structure & Function

Article Title: Somatostatin receptors (SSTR1-5) on inhibitory interneurons in the barrel cortex

doi: 10.1007/s00429-019-02011-7

Figure Lengend Snippet: SSTRs localisation in the somatosensory cortex of mice. High magnification confocal pictures of coronal sections presenting localisation of SSTR1-5 in the mouse somatosensory cortex. a SSTR1 labelled clearly the pyramidal-like cell bodies and apical dendrites. b Lack of co-localisation of SSTR2s and PV INs in the mouse primary somatosensory cortex. Yellow lines represent two orthogonal sections of a z-series showing the distribution of green and red fluorescence in the tissue. c SSTR3 immunoreactivity was found in cell bodies and neuropil. d, e Immunofluorescent signal for SSTR4 and SSTR5 was found mainly in the neuropil and much less visible in cell bodies. Scale bar for all pictures: 10 μm

Article Snippet: The primary antibodies used in this study were as follows: rabbit anti-SSTR1 polyclonal antibody (1:400, catalogue no. ASR-001, Alomone Labs), rabbit anti-SSTR2 monoclonal antibody (1:2000, catalogue no. ab134152, Abcam), rabbit anti-SSTR3 polyclonal antibody (1:300, catalogue no. ASR-003, Alomone Labs), rabbit anti-SSTR4 polyclonal antibody (1:1000, catalogue no. ab28578, Abcam), and rabbit anti-SSTR5 polyclonal antibody (1:1000, catalogue no. ASR-005, Alomone Labs).

Techniques: Confocal, Fluorescence, Tissue, Immunofluorescence

Journal: Brain Structure & Function

Article Title: Somatostatin receptors (SSTR1-5) on inhibitory interneurons in the barrel cortex

doi: 10.1007/s00429-019-02011-7

Figure Lengend Snippet: Two types of SSTRs distribution in the mouse barrel field. Tangential brain sections through L4 of the mouse barrel cortex. Type A distribution is characterised by a high concentration of immunoreactivity in the barrel walls (sides + septa). This type is represented by SSTR1 and SSTR2. Type B distribution is characterised by a homogeneous intensity of immunoreactivity in the barrel hollows. This type is represented by SSTR3, SSTR4, and SSTR5. Scale bar: 100 μm. Nuclear DAPI staining is visible in blue

Article Snippet: The primary antibodies used in this study were as follows: rabbit anti-SSTR1 polyclonal antibody (1:400, catalogue no. ASR-001, Alomone Labs), rabbit anti-SSTR2 monoclonal antibody (1:2000, catalogue no. ab134152, Abcam), rabbit anti-SSTR3 polyclonal antibody (1:300, catalogue no. ASR-003, Alomone Labs), rabbit anti-SSTR4 polyclonal antibody (1:1000, catalogue no. ab28578, Abcam), and rabbit anti-SSTR5 polyclonal antibody (1:1000, catalogue no. ASR-005, Alomone Labs).

Techniques: Concentration Assay, Staining, Immunofluorescence

Journal: Brain Structure & Function

Article Title: Somatostatin receptors (SSTR1-5) on inhibitory interneurons in the barrel cortex

doi: 10.1007/s00429-019-02011-7

Figure Lengend Snippet: Co-localisation of SSTRs with PV containing interneurons in the mouse primary somatosensory cortex. Coronal sections demonstrating the co-localisation of particular SSTRs (green immunofluorescence) with PV containing interneurons (red tdTomato fluorescence); DAPI staining shows the nuclei (blue). Scale bar: 100 μm

Article Snippet: The primary antibodies used in this study were as follows: rabbit anti-SSTR1 polyclonal antibody (1:400, catalogue no. ASR-001, Alomone Labs), rabbit anti-SSTR2 monoclonal antibody (1:2000, catalogue no. ab134152, Abcam), rabbit anti-SSTR3 polyclonal antibody (1:300, catalogue no. ASR-003, Alomone Labs), rabbit anti-SSTR4 polyclonal antibody (1:1000, catalogue no. ab28578, Abcam), and rabbit anti-SSTR5 polyclonal antibody (1:1000, catalogue no. ASR-005, Alomone Labs).

Techniques: Immunofluorescence, Fluorescence, Staining

Journal: Brain Structure & Function

Article Title: Somatostatin receptors (SSTR1-5) on inhibitory interneurons in the barrel cortex

doi: 10.1007/s00429-019-02011-7

Figure Lengend Snippet: Quantitative assessment of co-localisation of SSTRs with PV, SST, and VIP containing INs in the particular cortical layers of the mouse somatosensory cortex. Graphs represent the percentage (mean + SD) of all labelled interneurons of each particular type that exhibited co-localisation with SSTRs in different cortical layers. Top panel: SSTR1-5 + PV INs, Middle panel: SSTR1-5 + SST INs, Bottom panel: SSTR1-5 + VIP INs. Kruskal–Wallis test, * p ≤ 0.05, # 0.05 < p < 0.051; n = 3

Article Snippet: The primary antibodies used in this study were as follows: rabbit anti-SSTR1 polyclonal antibody (1:400, catalogue no. ASR-001, Alomone Labs), rabbit anti-SSTR2 monoclonal antibody (1:2000, catalogue no. ab134152, Abcam), rabbit anti-SSTR3 polyclonal antibody (1:300, catalogue no. ASR-003, Alomone Labs), rabbit anti-SSTR4 polyclonal antibody (1:1000, catalogue no. ab28578, Abcam), and rabbit anti-SSTR5 polyclonal antibody (1:1000, catalogue no. ASR-005, Alomone Labs).

Techniques:

Journal: Brain Structure & Function

Article Title: Somatostatin receptors (SSTR1-5) on inhibitory interneurons in the barrel cortex

doi: 10.1007/s00429-019-02011-7

Figure Lengend Snippet: Co-localisation of SSTRs with SST containing interneurons in the mouse primary somatosensory cortex. Coronal sections demonstrating the co-localisation of particular SSTRs (green immunofluorescence) with SST containing interneurons (red tdTomato fluorescence); DAPI staining shows the nuclei (blue). Scale bar 100 μm

Article Snippet: The primary antibodies used in this study were as follows: rabbit anti-SSTR1 polyclonal antibody (1:400, catalogue no. ASR-001, Alomone Labs), rabbit anti-SSTR2 monoclonal antibody (1:2000, catalogue no. ab134152, Abcam), rabbit anti-SSTR3 polyclonal antibody (1:300, catalogue no. ASR-003, Alomone Labs), rabbit anti-SSTR4 polyclonal antibody (1:1000, catalogue no. ab28578, Abcam), and rabbit anti-SSTR5 polyclonal antibody (1:1000, catalogue no. ASR-005, Alomone Labs).

Techniques: Immunofluorescence, Fluorescence, Staining

Journal: Brain Structure & Function

Article Title: Somatostatin receptors (SSTR1-5) on inhibitory interneurons in the barrel cortex

doi: 10.1007/s00429-019-02011-7

Figure Lengend Snippet: Co-localisation of SSTRs with VIP containing interneurons in the mouse primary somatosensory cortex. Coronal sections demonstrating the co-localisation of particular SSTRs (green immunofluorescence) with VIP containing interneurons (red tdTomato fluorescence); DAPI staining shows the nuclei (blue). Scale bar 100 μm

Article Snippet: The primary antibodies used in this study were as follows: rabbit anti-SSTR1 polyclonal antibody (1:400, catalogue no. ASR-001, Alomone Labs), rabbit anti-SSTR2 monoclonal antibody (1:2000, catalogue no. ab134152, Abcam), rabbit anti-SSTR3 polyclonal antibody (1:300, catalogue no. ASR-003, Alomone Labs), rabbit anti-SSTR4 polyclonal antibody (1:1000, catalogue no. ab28578, Abcam), and rabbit anti-SSTR5 polyclonal antibody (1:1000, catalogue no. ASR-005, Alomone Labs).

Techniques: Immunofluorescence, Fluorescence, Staining

Journal: bioRxiv

Article Title: Lineage transcription factors co-regulate subtype-specific genes providing a roadmap for systematic identification of small cell lung cancer vulnerabilities

doi: 10.1101/2020.08.13.249029

Figure Lengend Snippet: (A) RNA-seq (log2 FPKM) showing expression of SCN3A and KCNB2 , in SCLC tumors (T) or cell lines (C) in the specified SCLC subtype or NSCLC. The line indicates the mean, and the asterisks indicate p-values for each sample compared to SCLC-A tumors. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001. (B) UCSC Genome Browser tracks showing ASCL1, NKX2-1, PROX1 and H3K27ac ChIP-seq signals in NCI-H2107 cells around the genes encoding SCN3A and KCNB2 . Black bars above the tracks indicate computationally called binding sites. (C-E) Immunoblots showing SCN3A and KCNB2 protein in NCI-H2107 cells, 72 hours post-transfection with control siRNAs or siRNA targeted against either ASCL1, NKX2-1 , or PROX1 (C), or SCN3A (D) or KCNB2 (E). (F) WST-1 assay for cell viability from cells from (D,E). Each data point represents a biological replicate, and error bars indicate SEM. ANOVA with Bonferroni’s multiple comparisons test was used to determine significant differences relative to control. ns; not significant. (G) Quantification of colony formation assays in soft agar using varying doses of KCNB2 inhibitor Quinine shows increased sensitivity of SCLC-A NCI-H889, compared to SCLC-N NCI-H524, and SCLC-P NCI-H526. (H) Histograms showing SCN3A+ cells are detected in live SCLC-A but not SCLC-N or SCLC-P cell cultures using an SCN3A extracellular domain specific antibody (red). The background fluorescence with the secondary antibody only is shown (gray). (I,J) RT-qPCR for SCN3A (I) and ASCL1 (J) mRNA from FACs isolated SCN3A+ (red) or SCN3A-(black) cells from mixtures of SCLC-A with SCLC-P or – N. Each data point represents a biological sample, error bars = SD around mean, unpaired t-test, * p<0.05, ** p<0.01.

Article Snippet: Cells were incubated with SCN3A antibody (Alomone Labs, Anti-SCN3A (Nav1.3) (extracellular), Cat #: ASC-023, 4.25 μg/ml) for 30 min on ice, washed 3 times with ice cold FACs buffer, incubated with Alexa Fluor 568-conjugated secondary antibody (Thermofisher, Catalog # A10042, 1:400) for 30 min on ice, washed 3 times as above, and resuspended in ice cold FACs buffer.

Techniques: RNA Sequencing, Expressing, ChIP-sequencing, Binding Assay, Western Blot, Transfection, Control, WST-1 Assay, Colony Assay, Sensitivity, Fluorescence, Quantitative RT-PCR, Isolation, Chromatin Immunoprecipitation, ChIP-chip, FACS, Knockdown

Journal: Advanced Science

Article Title: Interior Decorative VOCs Elevate T Cell‐Mediated Obstructive Lung Disease Risks via Osteogenesis‐Driven Lymphoid‐Biased Hematopoiesis

doi: 10.1002/advs.202512663

Figure Lengend Snippet: VOCs exposure enhances T cell maturation and their recruitment to lung tissues. Mice were exposed to CON or VOCs for 8‐week using the whole‐body inhalation exposure model. A) Representative images of H&E‐stained thymic sections. Black dashed lines indicate the cortical thickness of thymus. Scale bar: 100 µm. Immunohistochemical analysis for B) CD3 and C) CD8 in thymic sections, and the percentage of positive area. Yellow arrows indicate the positive expression. Scale bar: 40 µm. D) Representative flow cytometric dot plots and quantitative analysis of E) CD4 + T and F) CD8 + T cells in the PB. G) Immunostaining for the colocalization of CXCR3 (red) and CD8 (purple) in lung sections, and the corresponding quantitative analysis. Yellow arrows indicate CXCR3 colocalization with CD8. Scale bar: 20 µm. H) Immunostaining for the colocalization of CCR5 (green) and CD8 (purple) in lung sections, and the corresponding quantitative analysis. Yellow arrows indicate CCR5 colocalization with CD8. Scale bar: 20 µm. I) Quantitative analysis of CXCL9, CXCL10, and CXCL11 in the PB by ELISA assay. Protein expressions of CXCL10 in J) BALF and K) LI. L) A schematic representation summarizing the T cell maturation and recruitment to lung tissue. Statistical analysis was performed using two‐tailed unpaired t ‐test (A–C and E–K). * p < 0.05 and ** p < 0.01. Data are expressed as mean ± SEM ( n ≥ 3).

Article Snippet: Similarly, mouse lung sections were incubated with primary antibodies against CXCR3 (Bosterbio, China), CCR5 (Servicebio, China), and CD8 (Servicebio, China) at 4 °C overnight, followed by the application of secondary antibodies.

Techniques: Staining, Immunohistochemical staining, Expressing, Immunostaining, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: bioRxiv

Article Title: Increased expression of developmental Nav1.3 promotes hippocampal CA3 hyperexcitability in early-stage 5xFAD mice

doi: 10.64898/2026.07.28.741298

Figure Lengend Snippet: a) Representative confocal images of dorsal hippocampus CA3 showing c-Fos (green, top), Nav1.3 + (red, middle), and merged channels (yellow, bottom) for WT and 5xFAD at 3 months (left) and 7 months (right). The inset on the top-left panel indicates the CA3 subregion delineated using the Allen Brain Atlas where c-Fos/Nav1.3 were quantified. Scale bar = 400 μm. b) Quantification of neuronal activation in CA3, shown as c-Fos + cells per 1000 μm 2 . c-Fos expression was higher in 3-month 5xFAD mice than in 3-month WT mice or 7-month 5xFAD mice (two-way ANOVA genotype x age interaction: F(1, 8) =12.0, p = 0.009; post-hoc Sidak: 5xFAD 3 months vs WT 3 months, p =0.015; 5xFAD 3 months vs 5xFAD 7 months, p = 0.049). c) Quantification of Nav1.3/c-Fos colocalization, shown as the percentage of c-Fos + neurons that are Nav1.3 +. The genotype x age interaction was not significant for colocalization, but there was a significant main effect of genotype, with higher Nav1.3/c-Fos co-expression at 3 months in 5xFAD mice vs age-matched WT controls (F(1, 8) = 11.8, p = 0.009); post-hoc Sidak’s comparisons indicate increased colocalization in 3-month 5xFAD mice relative to 3-month WT (p = 0.035). No other comparisons reached significance. Data are mean ± SD; n = 3 mice per group, 9 fields analyzed per mouse.

Article Snippet: An anti-Nav1.3 antibody (rabbit polyclonal; Alomone Labs #ASC-004; Jerusalem, Israel) was used, directed against an epitope in the C-terminal region of Nav1.3.

Techniques: Confocal, Activation Assay, Expressing, Immunofluorescence

Journal: bioRxiv

Article Title: Increased expression of developmental Nav1.3 promotes hippocampal CA3 hyperexcitability in early-stage 5xFAD mice

doi: 10.64898/2026.07.28.741298

Figure Lengend Snippet: a) Schematic of mossy fiber terminal (red) contact (red arrowhead, right) on CA3 pyramidal cell dendritic spine (light blue). Inset (left) shows an enlarged representation of mossy terminal-CA3 spine contact b) Representative pre-embedding immuno-electron micrographs showing Nav1.3 in a mossy-fiber terminal (top, presynaptic) and a CA3 dendritic spine (bottom, postsynaptic). Purple arrowheads indicate silver-intensified immunogold (SIG) labeling for Nav1.3. Scale bar = 400 nm. c) Quantification of Nav1.3 SIG particles from 50 MFTs in WT and 5xFAD mice at 3 and 7 months. Data are mean ± SD; n = 3 mice per group. Two-way ANOVA revealed a significant genotype x age interaction (F(1, 8) = 7.68, p = 0.024). Post-hoc Sidak comparisons showed increased Nav1.3 SIG particles in MFTs from 3-month 5xFAD mice compared with 3-month WT mice (p = 0.007); no other pairwise comparisons reached significance. d) Quantification of postsynaptic Nav1.3 SIG particles per 50 CA3 spines per mouse per condition. Two-way ANOVA showed no significant effect of genotype p = 0.42, age p = 0.96, or interaction p = 0.58; n= 3 mice per group.

Article Snippet: An anti-Nav1.3 antibody (rabbit polyclonal; Alomone Labs #ASC-004; Jerusalem, Israel) was used, directed against an epitope in the C-terminal region of Nav1.3.

Techniques: Labeling, Transmission Electron Microscopy

Journal: bioRxiv

Article Title: Increased expression of developmental Nav1.3 promotes hippocampal CA3 hyperexcitability in early-stage 5xFAD mice

doi: 10.64898/2026.07.28.741298

Figure Lengend Snippet: (a-c) Representative confocal images showing GCaMP6s expression (green, a), Nav1.3 immunolabeling (red, b) and merged channel (yellow, c) in dorsal CA3 following viral delivery. Arrowheads indicate examples of GCaMP6s + neurons with reduced Nav1.3 labeling after Nav1.3-shRNA; panel C also shows the implanted fiber tract (dashed outline) used for photometry. Scale bar = 400 μm d) Quantification of CA3 calcium activity (AUC/1000 frames) in 3-month-old WT, 5xFAD, 5xFAD+scrambled-shRNA, and 5xFAD+Nav1.3-shRNA mice. 3-month WT and 5xFAD data from are replotted for comparison. Nav1.3 knockdown significantly reduced CA3 activity relative to both untreated 5xFAD and 5xFAD+scrambled controls. Data are mean ± SD; n = 6-7 mice per group. One-way ANOVA was used to assess treatment effects at 3 months (F (3, 21) = 5.63, p = 0.005), followed by post-hoc tests (Tukey) for pairwise comparisons: 5xFAD vs 5xFAD+Nav1.3-shRNA, p = 0.030; 5xFAD+scrambled vs 5xFAD+Nav1.3-shRNA, p = 0.032; WT vs 5xFAD, p = 0.047. WT vs 5xFAD+Nav1.3-shRNA, p = 0.987. WT and 5xFAD here are the 3-month subset re-analyzed by one-way ANOVA, so the WT and 5xFAD p-value differs from the full two-way model in ).

Article Snippet: An anti-Nav1.3 antibody (rabbit polyclonal; Alomone Labs #ASC-004; Jerusalem, Israel) was used, directed against an epitope in the C-terminal region of Nav1.3.

Techniques: Confocal, Expressing, Immunolabeling, Labeling, shRNA, Activity Assay, Comparison, Knockdown, Immunofluorescence

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