Journal: Journal of translational medicine

Article Title: Apelin-13 attenuates optic nerve damage in glaucomatous mice by regulating glucose metabolism.

doi: 10.1186/s12967-025-06212-z

Figure Lengend Snippet: Fig. 4 Role of exogenous [Pyr1]-apelin-13 in glucose metabolism in retinal and brain neuronal cells. (A) GLUT1 expression in visual cortex neurons was elevated in the PECE group compared with the NC group, whereas it was lower in the Apelin-13 group than that in the PECE group. (B) The trend of GLUT3 expression in visual cortex neurons was consistent with that of GLUT1 expression. (C) G6PD expression in visual cortex neurons was lower in the PECE group than in the NC group, whereas G6PD expression was elevated in the Apelin-13 group compared with that in the PECE group. D a GLUT1, GLUT3, and G6PD expression in the retina. (D) b GLUT1 expression was decreased in the PECE group compared with the NC group and increased in the Apelin-13 group compared with that in the PECE group. (D) c GLUT3 expression was elevated in the PECE group compared with that in the NC group and decreased in the Apelin-13 group compared with the PECE group. (D) d G6PD expression was lower in the PECE group than in the NC group and increased in the Apelin-13 group compared with the PECE group. (E a GLUT1, GLUT3, and G6PD expression in the brain. (E) b c d Trend of GLUT1, GLUT3, and G6PD expression in the brain is consistent with that in the retinal tissues. *P < 0.05, **P < 0.01, ***P < 0.001. GLUT: glucose transporter; G6PD: glucose- 6-phosphate dehydrogenase; IOP: intraocular pressure; NC: normal control; PECE: PEG-PCL-PEG; RGCs: retinal ganglion cells

Article Snippet: The primary antibodies used were Iba1(1:200, anti-Iba1 antibody, Wako, 019-19741, Japan), NeuN (1:200, anti-NeuN antibody, Cell Signaling Technology, 94403 S, USA), GLUT1 (1:100, Novus, NB11039113, USA), GLUT3 (1:100, Novus, NBP3-12885, USA), and glucose-6-phosphate dehydrogenase (G6PD) (1:100, GeneTex, GTX101218, USA), followed by secondary antibodies (1:500, Bioworld, China).

Techniques: Expressing, Control

Journal: Journal of translational medicine

Article Title: Apelin-13 attenuates optic nerve damage in glaucomatous mice by regulating glucose metabolism.

doi: 10.1186/s12967-025-06212-z

Figure Lengend Snippet: Fig. 6 Effects of Apelin-13 on oxidative stress and the protein regulation and the PI3K/Akt pathway in retinal and brain neural tissue cells can be reversed by LY294002. (A) a NADPH levels in the retinal tissue were lower in the Apelin-13 group than in the PECE group, whereas they were higher in the LY294002 group than in the Apelin-13 group. (A) b NADPH levels in the brain tissues changed in the same pattern as those in the retinal tissues. (B) a ROS levels in the retinal tissues were lower in the Apelin-13 group than in the PECE group and higher in the LY294002 group. (B) b Trend of ROS levels in the brain tissues was the same as that in the retinal tissues. (C) The p-PI3K/PI3K and p-Akt/Akt ratios in the retinal tissues of mice were significantly higher in the Apelin-13 group than in the PECE group, whereas these ratios were reduced in the LY294002 group. (D) Alterations in p-PI3K/PI3K and p-Akt/Akt ratios in the brain tissues followed the same trend as observed in the retinal tissues. (E) a GLUT1, GLUT3, and G6PD expression in the retina. (E) b GLUT1 expression reduced in the LY294002 group compared with the Apelin-13 group. (E) c GLUT3 expression was increased in the LY294002 group compared with the Apelin-13 group. (E) d G6PD expression was reduced in the LY294002 group compared with the Apelin-13 group. (F) a GLUT1, GLUT3, and G6PD expres sion in the brain. (F b, c, d Trend of GLUT1, GLUT3, and G6PD expression in the brain was consistent with that in the retinal tissues. *P < 0.05, **P < 0.01, ***P < 0.001. Akt: protein kinase B; IOP: intraocular pressure; PI3K: phosphatidylinositol 3-kinase; RGCs: retinal ganglion cells; ROS: reactive oxygen species

Article Snippet: The primary antibodies used were Iba1(1:200, anti-Iba1 antibody, Wako, 019-19741, Japan), NeuN (1:200, anti-NeuN antibody, Cell Signaling Technology, 94403 S, USA), GLUT1 (1:100, Novus, NB11039113, USA), GLUT3 (1:100, Novus, NBP3-12885, USA), and glucose-6-phosphate dehydrogenase (G6PD) (1:100, GeneTex, GTX101218, USA), followed by secondary antibodies (1:500, Bioworld, China).

Techniques: Expressing

Journal: bioRxiv

Article Title: Endocrine-metabolic decoupling drives stress vulnerability in dystrophin deficiency

doi: 10.64898/2026.02.22.707291

Figure Lengend Snippet: a , Area under the curve (AUC) for blood glucose concentrations following a 5-h fast and intraperitoneal injection of insulin (0.5 IU kg⁻¹; insulin tolerance test (ITT)) measured at rest (non-stressed) or during a 2-h tube-restraint (stressed). b , Relative frequency distribution of pancreatic islet area from isolated islets. c , Mean pancreatic islet area. d-f , Representative immunoblot (d) and quantification of glucose transporter 3 (GLUT3) protein abundance in triceps (e) and brain (f) extracts, normalised to total protein and expressed relative to WT. g , [¹⁸F] fluorodeoxyglucose (¹⁸F-FDG) standardised uptake values (SUV) in templated brain regions from mice treated with exogeneous insulin prior to stress or non-stress conditions. Data were analysed using repeated-measures ANOVA or linear mixed-effects models with Bonferroni post-hoc adjustments, or One-way ANOVA or Kruskal-Wallis with Dunn’s post-hoc testing. n =6-8 per group. Data are mean ± SEM. * P < 0.05.

Article Snippet: After blocking in 5% BSA or skim milk in TBST for 1 h, membranes were incubated overnight at 4°C with primary antibodies for GLUT3 (ProteinTech, 1:500) or GLUT4 (Thermo Fisher, 1:500).

Techniques: Injection, Isolation, Western Blot, Quantitative Proteomics

Journal: The Journal of Neuroscience

Article Title: Structural and Functional Rescue of Chronic Metabolically Stressed Optic Nerves through Respiration

doi: 10.1523/jneurosci.3652-17.2018

Figure Lengend Snippet: Figure 1. Glycogen and glial glucose transporters decreased with glaucoma pathology. 905 A, Glycogen analysis in 3, 6, and 10-month DBA/2J (D2) and control DBA/2J-Gpnmb+ (D2G) optic nerve (ON), 906 (n=6 ON per group). B and F, Capillary electrophoresis of GLUT1 B, and GLUT3 F, protein in 3, 6, and 10-month 907 D2 and D2G ON normalized to total protein then to 3m D2G protein levels (n=8 ON per group). See Figure 1-1. 908 C, Distribution of GLUT1 in ON of 3m D2, and 10m D2G and D2 optic nerves immunolabeled with GFAP 909 (green). Arrows indicate colocalization of GLUT1 and GFAP, (n=3 sections per ON, 6 ON per group). D and E, 910 Glut1 and Glut3 mRNA levels in 3, 6, and 10-month D2 and D2G ON, normalized to Hprt mRNA then to 3m 911 D2G mRNA (n=5-7 ON per group). See Figure 1-1. G, Distribution of GLUT3 in ON stained with Fluoromyelin. 912 Arrows indicate colocalization of GLUT3 with Fluoromyelin, (n=3 sections per ON, 6 ON per group). H and I, 913 Percent of mean fluorescence intensity in the region of interest (ROI) for C, GLUT1 and G, GLUT3. All values are 914 presented as mean ± SEM, one-way ANOVA and Tukey’s post-hoc test. A, F(5, 30) =18.59, **p = 0.0022, ***p 915 = 0.00011; B, F(5, 42) = 6.463, **p = 0.0077; D, F(5, 30) = 1.906, **p = 0.001; E, F(5, 37) = 15.58, **p= 0.0023, 916 ***p =0.0001; H, F(2, 51) = 210.4, ***p = 0.0001. Scale bar=20μm for C and G. 917 918 Figure 2. Monocarboxylate transporters downregulated with glaucoma pathology. 919 A, Mct1 mRNA levels in 3, 6, and 10-month D2 and D2G ON, normalized to Hprt mRNA then to 3m D2G mRNA 920 (n=5-13 ON per group). See Figure 2-1. B, D and E, MCT1, MCT2 and MCT4 protein levels in 3, 6 and 10-month 921 D2G and D2 ON, normalized to total protein and then to 3m D2G protein levels (n=8 ON per group). C, F and G, 922 Distribution of MCT1 C, MCT2 F, and MCT4 G, in 3m D2, and 10m D2G and D2 ON stained with Fluoromyelin 923 (green) or immunolabeled with GFAP (green). Arrows indicate colocalization (n=3 sections per ON, 6 ON per 924 group). H, Distribution of MCT2 in human control and glaucoma patient ON, immunolabeled for β-tubulin 925 (green) and stained with DAPI (blue). Arrows indicate colocalization (n=4 sections per ON, 2 ON per group). 926 See Figure 2-1. I-L, Percent of mean fluorescence intensity in the ROI for I, MCT1, J, MCT2, K, MCT4 D2 and 927 D2G ON, and L MCT2, human ON. M, Quantification of total number of RGC axons in 6m D2, 10m D2G and 928 10m D2 ON. All values are presented as mean ± SEM, one-way ANOVA and Tukey’s post-hoc test. A, F(5, 47) = 929 4.579, *p = 0.0141; B, F(5, 38) = 6.553, **p = 0.0076; D, F(5, 42) = 16.14, *p = 0.0054, **p = 0.0013; E, F(5, 42) 930 = 4.734, *p= 0.0348; I, F(2, 51) = 173.2, ***p = 0.0001; J, F(2, 51) = 402.1, ***p = 0.0001; K, F(2, 51) = 192.6, 931 ***p = 0.0001; L, t(6) = 9.317, ***p = 0.0001, two-tailed unpaired t-test). Scale bar=20μm applies to C, F, G, 932 and H. 933 934 935 Figure 3. Low lactate accompanies AMPK activation and limits mitochondrial biogenesis and metabolic 936 cofactor pools. 937 A, L-lactate levels in 3, 6 and 10-month D2G and D2 ON (n=8 ON per group). B, Ratio of phosphorylated-AMPK 938 (pAMPK) to AMPK protein in 3, 6 and 10-month D2G and D2 ON (n=8 ON per group). C-D, Phosphorylated 939 AMPK immunofluorescence (magenta) and GFAP (green) micrographs in C, human control and glaucoma ON 940 (n=4 sections per ON, 2 ON per group); and D, 3m D2, and 10m D2G and D2 mice (n= 3 sections per ON, 6 ON 941 per group). Arrows indicate colocalization of pAMPK and GFAP. E-F, Percent of mean fluorescence intensity in 942 the region of interest (ROI) for pAMPK in E, 3m D2, and 10m D2G and D2 mice, and F, human. G-I, Analyses of 943 NAD+/NADH, creatine kinase (CK) activity and PGC1-α levels in 3, 6 and 10-month D2G and D2 mice. G, NAD+ 944 normalized to NADH levels (n=6 ON per group). See Figure 3-1. H, Creatine kinase activity normalized to total 945 protein (n=6 ON per group). I, PGC1-α protein levels normalized to total protein levels and then to 3m D2G 946 protein levels (n=8 ON per group). All values are presented as mean ± SEM, one-way ANOVA and Tukey’s post- 947 hoc test. A, F(5, 42) = 22.04, *p = 0.0124; B, F(5, 42) = 35.51, **p = 0.0032, ***p = 0.0001; E, F(2, 45) = 208.4, 948

Article Snippet: List of antibodies used for IHC and Capillary Electrophoresis analyses 1103 Antibody Dilution RRID Company Host AKT1 1:50 for WES AB_329827 Cell Signaling Technology Rabbit AMPK α1 2B7 1:200 for WES AB_2721834 Novus Biologicals Mouse BDNF 1:100 for IHC AB_630940 Santa Cruz Biotechnology Rabbit Brn3a 1:50 for IHC AB_626765 Santa Cruz Biotechnology Mouse βIII-tubulin 1:1000 for IHC AB_107216 Abcam Chicken GLUT1 1:200 for IHC, 1:50 for WES AB_790014 Novus Biologicals Rabbit GLUT3 1:25 for IHC and WES AB_2191306 R&D Systems Mouse Glutamine Synthetase 1:50 for WES AB_1127501 Santa Cruz Biotechnology Mouse GFAP 1:250 for IHC AB_10917109 EMD Millipore Mouse GFAP 1:500 for IHC AB_10013382 Dako-Agilent Rabbit MCT1 1:25 for IHC and WES AB_10841766 Santa Cruz Biotechnology Mouse MCT2 1:50 for WES AB_2721836 Aviva Systems Biology Rabbit 1:50 for WES AB_10855300 Bioss Rabbit MCT4 1:250 for IHC AB_2254765 Santa Cruz Biotechnology Goat 1:50 for WES AB_2189333 Santa Cruz Biotechnology Rabbit NRF2 1:25 for IHC AB_2618882 Developmental Studies Hybridoma Bank Mouse RBPMS 1:200 for IHC AB_10720427 GeneTex Inc. Rabbit PhosphoAKTSer 473 1:50 for WES AB_329825 Cell Signaling Technology Rabbit pAMPK α1Thr 172 1:100 for IHC, 1:50 for WES AB_11032916 Novus Biologicals Rabbit p70S6K 1:50 for WES AB_331676 Cell Signaling Technology Rabbit Phosphop70S6KThr 389 1:50 for WES AB_330944 Cell Signaling Technology Rabbit Phosphop70S6KSer 411 1:25 for WES AB_2182257 Santa Cruz Biotechnology Mouse SOD2 1:100 for IHC AB_2191814 Santa Cruz Biotechnology Mouse TFAM 1:50 for IHC AB_10610743 Santa Cruz Biotechnology Mouse 1104 48 48 1105 Table 2.

Techniques: Control, Electrophoresis, Immunolabeling, Staining, Fluorescence, Two Tailed Test, Activation Assay, Immunofluorescence, Activity Assay

Journal: Clinical epigenetics

Article Title: Epigenetic priming improves salvage chemotherapy in diffuse large B-cell lymphoma via endogenous retrovirus-induced cGAS-STING activation.

doi: 10.1186/s13148-023-01493-x

Figure Lengend Snippet: Fig. 7 A combination of vitamin C and 5-azacytidine could be a potential remedy for insufficient epigenetic priming. A IC50 values were calculated using cell viability measured on day 5. *p < 0.05, **p < 0.01 and ***p < 0.001 (two-way ANOVA). B, STING expression levels were measured using western blotting. C, D GLUT3 expression levels were measured using RNA-sequencing data and western blotting, GLUT3 is encoded by the SLC2A3 gene. E GLUT3 expression levels were measured using western blotting of empty vector-expressing and SLC2A3-overexpressing OCI-LY1 cell lines, OE: overexpression. F IC50 values were calculated using cell viability measured on day 5. *p < 0.05, **p < 0.01 and ***p < 0.001 (two-way ANOVA). G STING expression levels were measured using western blotting. H–J Subcutaneous tumor sizes, xenograft tumor images, and tumor weights of xenografts from empty vector-expressing or SLC2A3-overexpressing OCI-LY1 tumor-bearing models are shown. *p < 0.05, **p < 0.01 and ***p < 0.001 (t test). AZA: 5-azacytidine, VC: vitamin C, OE: overexpression

Article Snippet: A lentiviral vector for GLUT3 overexpression (lentiviral ORF clone of SLC2A3, mGFP tagged, CAT#: RC204430L4) and an empty vector (pLenti-C-mGFPP2A-Puro Lentiviral Gene Expression Vector, CAT#: PS100093) were purchased from Origene.

Techniques: Expressing, Western Blot, RNA Sequencing, Plasmid Preparation, Over Expression

Journal: eLife

Article Title: Insights into metabolic heterogeneity of colorectal cancer gained from fluorescence lifetime imaging

doi: 10.7554/eLife.94438

Figure Lengend Snippet: ( A ) Representative immunohistochemical images of GLUT3 expression. Scale bar = 50 μm (magnification x200) and 20 μm (magnification x630). ( B ) Representative immunohistochemical images of LDHA expression. Scale bar = 50 μm (magnification x200) and 20 μm (magnification x630). ( С ) Semi-quantitative evaluation of the expression level by staining intensity.

Article Snippet: Next, slides were incubated with primary polyclonal antibodies to Glucose Transporter 3 GLUT3 (E-AB-31557, Elabscience, China) or to Lactate dehydrogenase A LDHA (E-AB-19947, Elabscience, China) for 15 min. For the antibodies detection «Bond polymer refine detection system» (Leica Biosystems, UK) was used.

Techniques: Immunohistochemical staining, Expressing, Staining