Journal: bioRxiv

Article Title: Dual GLP-1 and GIP receptor agonist Tirzapetide plays an off-target role in the modulation of the β-adrenoceptors and glucose metabolism in hyperglycemic or senescent cardiac cells

doi: 10.1101/2025.04.09.648025

Figure Lengend Snippet: (A) The cellular resting level of Ca 2+ was imagined in all groups of H9c2 cells loaded with a Ca 2+ -sensitive fluorescence dye Fluo-3 AM. The representative images demonstrate the effect of TZPD (40 nM) on intracellular Ca 2+ distribution in resting cells with and without receptor antagonism for the SC group of cells (left) and HG group of cells (right). The average cellular Ca 2+ level in Fluo-3 AM loaded H9c2 cells was measured in all groups by the flow cytometer technique. Representative flow cytometry data are given in the left parts. The protein levels of phosphorylated PKA (pPKA) and PKA and their ratios (B), and phosphorylated GSK (pGSK) and GSK and their ratios (C) , compared to a reference protein GAPDH in the SC, SC+TZPD, SC+TZPD+ANT or HG, HG+TZPD, HG+TZPD+ANT comparison to the NC group. All shortens are the same as given in previous figures. Representative protein bands are given in the left parts of the bar graphs. Data are presented as means±SEM. * p <0.05 vs. NC group, γ p <0.05 vs. SC group, δ p <0.05 vs. HG group, Ψ p <0.05 vs. SC+ANT+TZPD group, τ p <0.05 vs. HG+ANT+TZPD group ( n =3-4 independent experiments).

Article Snippet: Following the transfer and blocking steps with 1% BSA in TBS-0.3% Tween, the membrane was incubated with the primer antibodies of either β1-AR (Thermo,), β2-AR (Origine, NM_012492), β3-AR (Saint Jones, STJ91728-200), GLP-1R (MedChem, HY-P81160) GIP-R (MedChem, HY-P10138), PKG (Thermo, PA3-031A), Glut4 (Medchem, HY-P80689), pPKA (Cell signaling, 4781), PKA (Cell signaling, 4782), pGSK (Santa Cruz, sc-373800), GSK (Santa Cruz, sc-377213), eNOS (Thermo, PA3-031A), alpha tubulin (Santa Cruz, sc-5286), and GAPDH (Santa Cruz, sc-137179) primer antibody to use as house-keeping control.

Techniques: Fluorescence, Flow Cytometry, Comparison

Journal: Stem cell research & therapy

Article Title: Therapeutic effect of adipose stromal vascular fraction spheroids for partial bladder outlet obstruction induced underactive bladder.

doi: 10.1186/s13287-022-02739-w

Figure Lengend Snippet: Fig. 8 Effects of ad-SVF spheroids on cell apoptosis and proliferation. A Representative images of TUNEL staining in each group. B Representative images of PCNA staining in each group. C, D Semi-quantitative analysis of TUNEL-positive cells and PCNA-positive cells was performed by using Image J software. E Representative western blot images of AKT, pAKT, GSK-3β, pGSK-3β, and GAPDH. F Relative abundance of AKT/GAPDH, pAKT/ GAPDH, GSK-3β/GAPDH and pGSK-3β/GAPDH were quantified according to western blots. Scale bar = 100 μm. *p < 0.05 (ad-SVF, ad-SVFsp vs Sham); #p < 0.05 (ad-SVFsp vs ad-SVF)

Article Snippet: The membranes were incubated with primary antibodies (bFGF (1:1000, Bioss), HGF (1:1000, Bioss), VEGFA (1:1000, Bioss), AKT (1:2000, Proteintech), pAKT (1:2000, Proteintech), GSK-3β(1:1000, Proteintech), pGSK-3β(1:2000, Proteintech), and GAPDH (1:2000, Proteintech)) overnight at 4 °C after blocked with 5% non-fat milk, then followed by a secondary horseradish peroxidase-conjugated antibody (anti-rabbit or anti-mouse (1:5000, Cell Signaling)) for 2 h at room temperature.

Techniques: TUNEL Assay, Staining, Software, Western Blot