Journal: Scientific Reports

Article Title: TCF7L2 involvement in estradiol- and progesterone-modulated islet and hepatic glucose homeostasis

doi: 10.1038/srep24859

Figure Lengend Snippet: MIN6 cells were plated at 5 × 10 5 cells per well in 6-well plates and exposed to a TCF7L2-specific short hairpin RNA (shTCF7L2) or a scrambled shRNA (shScr) for 72 h, then cultured for 24 h in the presence of 100 nM E 2 or 1 μM P 4 . ( A ) Western blots showing the TCF7L2 protein content after E 2 or P 4 treatment. ( B ) Viable cells. ( C,D ) Basal and stimulated insulin/proinsulin secretions (normalized to viable cell numbers). ( E,F ) Stimulatory indexes. ( G ) Proinsulin-to-insulin ratio. * P < 0.05 vs. sex hormone treatment control; # P < 0.05 shTCF7L2 vs. shScr.

Article Snippet: The antibodies previously used for Western blots were the anti-human TCF7L2 antibody (1:800; 13838-1-AP, Proteintech, USA), anti-human PEPCK antibody (1:1000; 14892-1-AP; Proteintech, USA), anti-human GLUT2 antibody (1:800; 20436-1-AP; Proteintech, USA), anti-human IRS2 antibody (1:1000; 20702-1-AP; Proteintech, USA), anti-human pAKT antibody (1:1000; 60072-1-Ig; Proteintech, USA), anti-human AKT antibody (1:1000; 10176-2-AP; Proteintech, USA), anti-human pGSK antibody (1:1000; 14850-1-AP; Proteintech, USA), anti-human GSK antibody (1:1000; 22104-1-AP; Proteintech, USA), anti-human pERK1/2 antibody (1:1000; 3441-100; BioVision, USA), anti-human ERK1/2 antibody (1:1000; 16443-1-AP; Proteintech, USA), and anti-human GAPDH antibody (1:1000; 10494-1-AP; Proteintech, USA).

Techniques: shRNA, Cell Culture, Western Blot, Control

Journal: Scientific Reports

Article Title: TCF7L2 involvement in estradiol- and progesterone-modulated islet and hepatic glucose homeostasis

doi: 10.1038/srep24859

Figure Lengend Snippet: MIN6 cells were plated at 5 × 10 5 cells per well in 6-well plates and exposed to high glucose concentration (33.3 mM) and transfected with TCF7L2-IRES2-EGFP (OE-TCF7L2) or a control vector (CV) for 72 h, then cultured for 24 h in the presence of 100 nM E 2 or 1 μM P 4 . ( A ) Western blots showing the TCF7L2 protein content after E 2 or P 4 treatment. ( B ) Viable cells. ( C , D ) Basal and stimulated insulin/proinsulin secretions (normalized to viable cell numbers). ( E,F ) Stimulatory indexes. ( G ) Proinsulin-to-insulin ratio. * P < 0.05 vs. sex hormone treatment control; # P < 0.05 OE-TCF7L2 vs. CV.

Article Snippet: The antibodies previously used for Western blots were the anti-human TCF7L2 antibody (1:800; 13838-1-AP, Proteintech, USA), anti-human PEPCK antibody (1:1000; 14892-1-AP; Proteintech, USA), anti-human GLUT2 antibody (1:800; 20436-1-AP; Proteintech, USA), anti-human IRS2 antibody (1:1000; 20702-1-AP; Proteintech, USA), anti-human pAKT antibody (1:1000; 60072-1-Ig; Proteintech, USA), anti-human AKT antibody (1:1000; 10176-2-AP; Proteintech, USA), anti-human pGSK antibody (1:1000; 14850-1-AP; Proteintech, USA), anti-human GSK antibody (1:1000; 22104-1-AP; Proteintech, USA), anti-human pERK1/2 antibody (1:1000; 3441-100; BioVision, USA), anti-human ERK1/2 antibody (1:1000; 16443-1-AP; Proteintech, USA), and anti-human GAPDH antibody (1:1000; 10494-1-AP; Proteintech, USA).

Techniques: Concentration Assay, Transfection, Control, Plasmid Preparation, Cell Culture, Western Blot

Journal: Scientific Reports

Article Title: TCF7L2 involvement in estradiol- and progesterone-modulated islet and hepatic glucose homeostasis

doi: 10.1038/srep24859

Figure Lengend Snippet: HepG2 cells (2.5 × 10 5 cells per well) were seeded in 6-well plates and exposed to a TCF7L2-specific short hairpin RNA (shTCF7L2) or a scrambled shRNA (shScr) for 72 h, or transfected with TCF7L2-IRES2-EGFP (OE-TCF7L2) or a control vector (CV) for 72 h, then cultured for 24 h in the presence of 100 nM E 2 or 1 μM P 4 . ( A,B ) 2-NBDG uptake. ( C,D ) Glucose production. ( E,F ) Western blot. * P < 0.05 vs. sex hormone treatment control; # P < 0.05 shTCF7L2 vs. shScr or OE-TCF7L2 vs. CV.

Article Snippet: The antibodies previously used for Western blots were the anti-human TCF7L2 antibody (1:800; 13838-1-AP, Proteintech, USA), anti-human PEPCK antibody (1:1000; 14892-1-AP; Proteintech, USA), anti-human GLUT2 antibody (1:800; 20436-1-AP; Proteintech, USA), anti-human IRS2 antibody (1:1000; 20702-1-AP; Proteintech, USA), anti-human pAKT antibody (1:1000; 60072-1-Ig; Proteintech, USA), anti-human AKT antibody (1:1000; 10176-2-AP; Proteintech, USA), anti-human pGSK antibody (1:1000; 14850-1-AP; Proteintech, USA), anti-human GSK antibody (1:1000; 22104-1-AP; Proteintech, USA), anti-human pERK1/2 antibody (1:1000; 3441-100; BioVision, USA), anti-human ERK1/2 antibody (1:1000; 16443-1-AP; Proteintech, USA), and anti-human GAPDH antibody (1:1000; 10494-1-AP; Proteintech, USA).

Techniques: shRNA, Transfection, Control, Plasmid Preparation, Cell Culture, Western Blot

Journal: Cell Death & Disease

Article Title: The Janus face of endogenous neuronal tPA: promoting self-protection and worsening the death of neighboring neurons

doi: 10.1038/s41419-024-06655-0

Figure Lengend Snippet: A Schematic experimental protocol showing the unilateral injection of 2 × 0.25 µL of 1.25 × 10 13 vp/mL of AAV9-pPlat-GFP in tPA flox −/− mice (WT, -0.3 mm AP, ±3.3 mm ML, –0.8 & 0.4 mm DV). B Representative images of immunohistochemical analyses showing that AAV9-pPlat-GFP does not transduce oligodendrocytes (magenta), microglia (red), astrocytes (gray), and blood vessels (cyan). Scale bar = 100 µm, ×20. C Representative images and corresponding quantifications of immunohistochemical analyses showing that AAV9-Plat -GFP transduces around one third of neurons (yellow; NeuN and Blue; Neurotrace). Scale bar = 100 µm, objective = ×40. D Quantification of the proportion of neurons GABA-expressing (a marker of inhibitory neurons) or tbr1 (a marker of excitatory neurons) among the population of GFP (tPA) positive neurons, in the transduced area. A white dashed arrow indicates the colocalization between the TBR1 marker and GFP, whereas an isolated white triangle denotes the colocalization between the GABA marker and GFP. Scale bar=100 µm, ×20.

Article Snippet: The following antibodies and/or tracers were used: mouse anti-NeuN (1:800, MAB377, Merck); rabbit anti-Green fluorescent protein (GFP, 1:1000, ab6556, abcam); goat anti-Green fluorescent protein (GFP, 1:1000, ab5450, abcam); chicken anti-Glial Fibrillary Acidic Protein (GFAP, 1:2000, ab4674, abcam); rabbit anti-Ionized calcium binding adapter molecule 1 (Iba-1, 1:1000, 019-19741, Fujifilm); rabbit anti- Gamma aminobutyric acid (GABA, 1:500, A2052, Sigma Aldrich); Rabbit anti-CD31 (1:1000, ab28364, abcam); NeuroTrace™ 435/455 Blue Fluorescent Nissl Stain (1:300, N21479, Invitrogen); rabbit anti- Oligodendrocyte transcription factor (Olig2, 1:3000, ab9610, Millipore); chicken anti-T-box brain transcription factor 1 (tbr1, 1:250, ab2261, Merck).

Techniques: Injection, Immunohistochemical staining, Expressing, Marker, Isolation

Journal: Cell Death & Disease

Article Title: The Janus face of endogenous neuronal tPA: promoting self-protection and worsening the death of neighboring neurons

doi: 10.1038/s41419-024-06655-0

Figure Lengend Snippet: A , B Representative images and quantification ( C ) of the percentage of tPA-expressing neurons (green), inhibitory tPA-expressing neurons (GABA + , red) and tPA-expressing neurons (red) and excitatory tPA-expressing neurons (tbr1 + , yellow) mortality 6-h post ischemia for WT and tPA-cKO Neu mice ( N = 5 for WT and 4 for tPA-cKO Neu mice; unpaired t -test; p < 0.05 = *). Scale bar = 100 µm, ×20.

Article Snippet: The following antibodies and/or tracers were used: mouse anti-NeuN (1:800, MAB377, Merck); rabbit anti-Green fluorescent protein (GFP, 1:1000, ab6556, abcam); goat anti-Green fluorescent protein (GFP, 1:1000, ab5450, abcam); chicken anti-Glial Fibrillary Acidic Protein (GFAP, 1:2000, ab4674, abcam); rabbit anti-Ionized calcium binding adapter molecule 1 (Iba-1, 1:1000, 019-19741, Fujifilm); rabbit anti- Gamma aminobutyric acid (GABA, 1:500, A2052, Sigma Aldrich); Rabbit anti-CD31 (1:1000, ab28364, abcam); NeuroTrace™ 435/455 Blue Fluorescent Nissl Stain (1:300, N21479, Invitrogen); rabbit anti- Oligodendrocyte transcription factor (Olig2, 1:3000, ab9610, Millipore); chicken anti-T-box brain transcription factor 1 (tbr1, 1:250, ab2261, Merck).

Techniques: Expressing