Journal: Cardiovascular Diabetology

Article Title: The dual GLP-1 and GIP receptor agonist tirzapetide provides an unintended interaction with the β-adrenoceptors and plays a role in glucose metabolism in hyperglycemic or senescent cardiac cells

doi: 10.1186/s12933-025-02828-z

Figure Lengend Snippet: Effect of the TZPD application on the cellular resting level of Ca 2+ and protein levels of parameters played roles in cellular Ca 2+ regulation via β-AR signaling. 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 ( A ). The representative flow cytometry data 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 and HG group of cells (left). The average cellular Ca 2+ level in Fluo-3 AM loaded H9c2 cells was measured in all groups by the flow cytometer technique (right). 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 shortenings 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. The comparisons between the two groups are performed by t-test. * 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 = 4 for flow cytometry experiments and n = 3 for western blotting 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, PA3-031 A), GLUT4 (Medchem, HY-P80689), pPKA (Cell signaling, 4781), PKA (Cell signaling, 4782), pGSK (Santa Cruz, sc-373800), GSK (Santa Cruz, sc-377213), eNOS (Thermo, PA3-031 A), 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, Western Blot