Journal: Frontiers in Veterinary Science

Article Title: Estrogen Regulates Glucose Metabolism in Cattle Neutrophils Through Autophagy

doi: 10.3389/fvets.2021.773514

Figure Lengend Snippet: E2 regulates glucose uptake and utilization by regulating the activities of HK and glucose transporters. PMNs were treated with LG, NG, HG, LG+EL, LG+EM, LG+EH, NG+EL, NG+EM, NG+EH, HG+EL, HG+EM and HG+EH for 6 h. (A–D) HK activity was measured by the biochemical method. (E–H) The expression of GLUT1, GLUT4 and SGLT1 were detected by WB, and β-actin was used as an internal control. The results are shown as the mean ± SD ( n = 3). The significance of the difference was analyzed using the t -test. * P < 0.05, ** P < 0.01.

Article Snippet: After blocking with 5% BSA for 2 h, the membranes were blotted with 1:700 diluted primary antibodies against GLUT1 (Abcam, MA, USA), GLUT4 (Abcam), SGLT1 (Cell Signaling Technology, MA, USA), Beclin1 (Abcam), ATG5 (Abcam), p62 (Abcam), LC3 (Cell Signaling Technology), β-actin (Absin), AMPK (Abcam), p-AMPK (PL Laboratories, USA), Bax (Abcam), or Bcl-2 (Abcam) at 4°C overnight.

Techniques: Activity Assay, Expressing, Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Sodium-dependent glucose transporter-1 as a novel immunological player in the intestinal mucosa.

doi: 10.4049/jimmunol.181.5.3126

Figure Lengend Snippet: FIGURE 2. SGLT-1 expression in IECs and siRNA-mediated silencing. A, mRNA expression of SGLT-1 in HT29, LCC-18, STC-1, and RAW 264.7. Lane 1, cDNA-containing sample; lane 2, water control; lane 3, RT control. B, HT29, after siRNA-mediated silencing of SGLT-1, were left untreated or treated with LPS with or without D-glucose pretreatment. Su- pernatants collected after 24 h were assayed for IL-8 production by ELISA. Data are the means (SD) of four independent experiments performed in triplicate. siR, siRNA. , p 0.001 siR Glu LPS vs Glu LPS.

Article Snippet: Briefly, HT-29 cells (3 105/well) seeded in six-well plates at 60–80% of confluence were washed in Optimem (Invitrogen) medium and then transfected with a pool of siRNA oligonucleotides targeting human SGLT-1 or a scrambled RNA duplex (Santa Cruz Biotechnology), at a final concentration of 100 nM.

Techniques: Expressing, Control, Enzyme-linked Immunosorbent Assay

Journal: Pharmaceuticals

Article Title: Dual-Target Compounds against Type 2 Diabetes Mellitus : Proof of Concept for Sodium Dependent Glucose Transporter (SGLT) and Glycogen Phosphorylase (GP) Inhibitors

doi: 10.3390/ph14040364

Figure Lengend Snippet: Inhibitory effects (IC 50 [nM]) of phlorizin and selected gliflozins against sodium dependent glucose cotransporter (SGLT2) (selectivity SGLT2/SGLT1).

Article Snippet: The membrane was incubated with anti-SGLT1 (anti-SLC5A1 TA324226 from Origene, Rockville, MD, USA), anti-SGLT2 (ab137207, Abcam, Cambridge, UK), or anti-GAPDH (ABS16, EMD Millipore, Rockville, MD, USA) in 1% BSA solution.

Techniques:

Journal: Pharmaceuticals

Article Title: Dual-Target Compounds against Type 2 Diabetes Mellitus : Proof of Concept for Sodium Dependent Glucose Transporter (SGLT) and Glycogen Phosphorylase (GP) Inhibitors

doi: 10.3390/ph14040364

Figure Lengend Snippet: Expression of SGLT1 ( A , B ) and SGLT2 ( C , D ) in transfected TSA201 cells and mouse kidney by Western blot. GAPDH was used to normalize for protein loading. The results were quantified by BIORAD Image Lab 5.2.1 software. (* p < 0.05 vs. TSA 201, n = 3 independent experiment per group). The kidney is a positive control to validate our SGLT1 and 2 antibodies. Full blot pictures are shown in .

Article Snippet: The membrane was incubated with anti-SGLT1 (anti-SLC5A1 TA324226 from Origene, Rockville, MD, USA), anti-SGLT2 (ab137207, Abcam, Cambridge, UK), or anti-GAPDH (ABS16, EMD Millipore, Rockville, MD, USA) in 1% BSA solution.

Techniques: Expressing, Transfection, Western Blot, Software, Positive Control

Journal: Pharmaceuticals

Article Title: Dual-Target Compounds against Type 2 Diabetes Mellitus : Proof of Concept for Sodium Dependent Glucose Transporter (SGLT) and Glycogen Phosphorylase (GP) Inhibitors

doi: 10.3390/ph14040364

Figure Lengend Snippet: Glucose uptake assay of control and SGLT1 ( A ) and SGLT2 ( B )-transfected TSA201 cells. Cells were treated with 2-NBDG (2-deoxy-2-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]-D-glucose) in the presence or absence of phlorizin. Glucose uptake was increased significantly in SGLT1 and SGLT2-transfected cells compared with control TSA201 cells. Phlorizin significantly decreased glucose uptake in both control and SGLT1 and SGLT2-transfected cells. ( n = 3 per group; * p < 0.05).

Article Snippet: The membrane was incubated with anti-SGLT1 (anti-SLC5A1 TA324226 from Origene, Rockville, MD, USA), anti-SGLT2 (ab137207, Abcam, Cambridge, UK), or anti-GAPDH (ABS16, EMD Millipore, Rockville, MD, USA) in 1% BSA solution.

Techniques: Transfection

Journal: Pharmaceuticals

Article Title: Dual-Target Compounds against Type 2 Diabetes Mellitus : Proof of Concept for Sodium Dependent Glucose Transporter (SGLT) and Glycogen Phosphorylase (GP) Inhibitors

doi: 10.3390/ph14040364

Figure Lengend Snippet: Inhibitory potencies (IC 50 [µM] a ) of the new and some previously prepared compounds against SGLT1, SGLT2, and rabbit muscle glycogen phosphorylase b (rmGPb).

Article Snippet: The membrane was incubated with anti-SGLT1 (anti-SLC5A1 TA324226 from Origene, Rockville, MD, USA), anti-SGLT2 (ab137207, Abcam, Cambridge, UK), or anti-GAPDH (ABS16, EMD Millipore, Rockville, MD, USA) in 1% BSA solution.

Techniques:

Journal: Pharmaceuticals

Article Title: Dual-Target Compounds against Type 2 Diabetes Mellitus : Proof of Concept for Sodium Dependent Glucose Transporter (SGLT) and Glycogen Phosphorylase (GP) Inhibitors

doi: 10.3390/ph14040364

Figure Lengend Snippet: Inhibitory potencies (IC 50 [µM] a ) of some aryl-substituted glucopyranosyl azole type glycogen phosphorylase inhibitors b against SGLT2 and SGLT1.

Article Snippet: The membrane was incubated with anti-SGLT1 (anti-SLC5A1 TA324226 from Origene, Rockville, MD, USA), anti-SGLT2 (ab137207, Abcam, Cambridge, UK), or anti-GAPDH (ABS16, EMD Millipore, Rockville, MD, USA) in 1% BSA solution.

Techniques:

Journal: Physiological Reports

Article Title: Acute tuft cell ablation in mice induces malabsorption and alterations in secretory and immune cell lineages in the small intestine

doi: 10.14814/phy2.70264

Figure Lengend Snippet: Primary antibody list.

Article Snippet: SGLT1 , Rabbit , Unconjugated , Novus , NBP2‐38748 , 1:200.

Techniques: Conjugation Assay

Journal: RSC Advances

Article Title: Synthesis, radiolabeling, and biological evaluation of methyl 6-deoxy-6-[ 18 F]fluoro-4-thio-α- d -maltotrioside as a positron emission tomography bacterial imaging agent †

doi: 10.1039/d5ra00693g

Figure Lengend Snippet: Immunohistochemistry on tissue around the skin pocket. Expression of SGLT1 and SGLT2 in the mock-up area was evaluated by immunohistochemistry. (a) SGLT1 (red) was expressed on fibroblasts (green, vimentin-positive cells), which exhibited an increase in the inflammatory tissue. (b) Similarly, SGLT2 (red) was expressed on fibroblasts (green). The scale bars represent 40 μm.

Article Snippet: Human SGLT1 (hSGLT1) and SGLT2 (hSGLT2) cDNA clones were obtained from R&D systems (RDC0654 and RDC0557, respectively).

Techniques: Immunohistochemistry, Expressing

Journal: RSC Advances

Article Title: Synthesis, radiolabeling, and biological evaluation of methyl 6-deoxy-6-[ 18 F]fluoro-4-thio-α- d -maltotrioside as a positron emission tomography bacterial imaging agent †

doi: 10.1039/d5ra00693g

Figure Lengend Snippet: Expression of SGLT1 and SGLT2 in the skin pocket from the control group and from the non-infectious inflammation group was quantified with qPCR. The expression of the target genes is indicated as the ratio to β-actin, a house keeping gene. Expression of SGLT1 in the non-infectious inflammation group was about 12 times that of the control group. Expression of SGLT2 in the non-infectious inflammation group was about 1.8 times the control group.

Article Snippet: Human SGLT1 (hSGLT1) and SGLT2 (hSGLT2) cDNA clones were obtained from R&D systems (RDC0654 and RDC0557, respectively).

Techniques: Expressing, Control, Gene Expression

Journal: RSC Advances

Article Title: Synthesis, radiolabeling, and biological evaluation of methyl 6-deoxy-6-[ 18 F]fluoro-4-thio-α- d -maltotrioside as a positron emission tomography bacterial imaging agent †

doi: 10.1039/d5ra00693g

Figure Lengend Snippet: Accumulation of [ 18 F]MFTMT and derivative of [ 18 F]MFTMT by SGLT1. (a) Stable expression of human SGLT1 (hSGLT1) or SGLT2 (hSGLT2) was established in CHO-K1 cells. CHO-K1-hSGLT1 and CHO-K1 hSGLT2 cells internalized 1-NBDG, a specific substrate for SGLT1 and SGLT2. Scale bars represent 100 μm. (b) Retention of [ 18 F]MFTMT and (c) retention of [ 18 F]MFTM by CHO-K1 cells overexpressing SGLTs. CHO-K1-hSGLT1, hSGLT2, and control cells were incubated with 20 μCi mL −1 of [ 18 F]MFTMT for 1 h, and the remaining radioactivity was evaluated. Some cells were treated with vehicle, mizagliflozin, a specific SGLT1 inhibitor, or dapagliflozin, a specific SGLT2 inhibitor. Retention of both [ 18 F]MFTMT and [18F]MFTM in CHO-K1-hSGLT1 cells were significantly higher compared to CHO-K1-hSGLT2 and control cells, and mizagliflozin significantly suppressed the retention of [ 18 F]MFTMT and [ 18 F]MFTM.

Article Snippet: Human SGLT1 (hSGLT1) and SGLT2 (hSGLT2) cDNA clones were obtained from R&D systems (RDC0654 and RDC0557, respectively).

Techniques: Expressing, Control, Incubation, Radioactivity

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Human Intestinal Enteroids With Inducible Neurogenin-3 Expression as a Novel Model of Gut Hormone Secretion

doi: 10.1016/j.jcmgh.2019.04.010

Figure Lengend Snippet: Antibodies Used in This Study

Article Snippet: Primary , Sodium-glucose transporter 1/SLC5A1 , Rabbit , Novus Biologicals (Centennial, CO) , NBP-238748 , 1:100.

Techniques:

Journal: Journal of ginseng research

Article Title: Ginsenoside Rd protects cerebral endothelial cells from oxygen-glucose deprivation/reoxygenation induced pyroptosis via inhibiting SLC5A1 mediated sodium influx.

doi: 10.1016/j.jgr.2022.05.006

Figure Lengend Snippet: Fig. 3. Ginsenoside Rd binds to SLC5A1 with a high affinity. A. Predicted docking protein of ginsenoside Rd, by using TargetNet. B. Molecular docking of between ginsenoside Rd and SLC5A1. The estimated thee binding positions and related residuals within SLC5A1 were provided in the bottom panel. C. Left: schematic of proposed interactions between SLC5A1 and glucose by one previous study [25]. X, XI, XII and XIII are four translocation domains of SLC5A1. The magenta ellipse indicates the aromatic stacking with the hydrophobic surface of the pyranose. Right: Three-dimensional model of the glucose binding domains of SLC5A1 [25]. The same helical domains in the left and right panels are labeled with the same color. Residues on the external surface are highlighted with yellow boxes in the right panel. D. SPR analysis of ginsenoside Rd binding to human recombinant SLC5A1 protein. E-F. Representative images (E) and quantitation (F) of CETSA of SLC5A1 with 30 mM ginsenoside Rd. n ¼ 3 independent experiments. **p < 0.01, ***p < 0.001.

Article Snippet: Human recombinant SLC5A1 protein (Novus Biologicals, Littleton, CO, USA) was captured on a CM5 chip via a standard amine coupling procedure.

Techniques: Binding Assay, Translocation Assay, Labeling, Recombinant, Quantitation Assay

Journal: Journal of ginseng research

Article Title: Ginsenoside Rd protects cerebral endothelial cells from oxygen-glucose deprivation/reoxygenation induced pyroptosis via inhibiting SLC5A1 mediated sodium influx.

doi: 10.1016/j.jgr.2022.05.006

Figure Lengend Snippet: Fig. 4. Ginsenoside Rd reduces OGD/R-induced sodium influx and potassium efflux in HBMECs. A. SLC5A1 mRNA expression in HBMECs after OGD/R stimulation, with or without treatment of ginsenoside Rd. B. SLC5A1 and SLC5A2 protein expression in HBMECs after OGD/R stimulation, with or without treatment of ginsenoside Rd. C-D. Relative intracellular Naþ (C) and Kþ (D) were detected in HBMECs after OGD(2 h)/R (1 h) stimulation, with or without the presence of ginsenoside Rd. E-F. Relative TEER values (E) and relative concentration of FITC-dextran (F) were calculated to assess the protective effect of phlorizin (25 and 50 mM) against OGD/R-induced damage of barrier function in different OGD/R time. G-J. Representative immunofluorescent images (G) and western blotting assays (HeJ) showing the expression of ZO-1 (green) and claudin-5 (red) in HBMECs after OGD(2 h)/R (12 h), with or without phlorizin treatment (50 mM). Scale bar ¼ 20 mm. Results were reported as mean ± SD (n ¼ 3). ***p < 0.001. n.s.: not significant.

Article Snippet: Human recombinant SLC5A1 protein (Novus Biologicals, Littleton, CO, USA) was captured on a CM5 chip via a standard amine coupling procedure.

Techniques: Expressing, Concentration Assay, Western Blot

Journal: Journal of ginseng research

Article Title: Ginsenoside Rd protects cerebral endothelial cells from oxygen-glucose deprivation/reoxygenation induced pyroptosis via inhibiting SLC5A1 mediated sodium influx.

doi: 10.1016/j.jgr.2022.05.006

Figure Lengend Snippet: Fig. 5. Inhibiting SLC5A1 suppresses OGD/R-activated NLRP3 inflammasome and pyroptosis in HBMECs. A-E. Representative blots (A) and quantitative analysis (BeE) of NLRP3, cleaved caspase-1, IL-1b and GSDMD-NT expression in HBMECs after OGD stimulation, with or without phlorizin treatment. F-G. Pyroptotic cells with the same treatment in panel A were assessed by caspase-1 and PI staining. Representative flow cytometric images (F) and quantitative data (G) of caspase-1þ/PI þ cell ratio were provided. Results were reported as mean ± SD (n ¼ 3). ***p < 0.001.

Article Snippet: Human recombinant SLC5A1 protein (Novus Biologicals, Littleton, CO, USA) was captured on a CM5 chip via a standard amine coupling procedure.

Techniques: Expressing, Staining