Journal: iScience

Article Title: PKA-mediated BMAL1 phosphorylation promotes β 1 -adrenoceptor autoantibody-induced cardiomyocyte death

doi: 10.1016/j.isci.2025.112786

Figure Lengend Snippet: β 1 -AA induces cytoplasmic accumulation of BMAL1 in H9c2 cells (A) Subcellular localization of BMAL1 was detected by immunofluorescence staining. BMAL1 staining (left), DAPI staining (middle) to visualize nuclei, and merged images (right). Scale bars: 15 μm. (B) BMAL1 levels in cytosolic and nuclear fractions at CT8 were detected by western blot, with GAPDH (cytoplasm) and histone H3 (nucleus) serving as markers. Data are represented as mean ± SEM.

Article Snippet: The following primary antibodies were used: BMAL1, phospho-BMAL1 (Ser42), Histone H3, phospho-PKA substrate (Cell Signaling Technology, USA); PRKACA (Immunoway, China); Tubulin (Proteintech, China) and GAPDH (Bioss, China).

Techniques: Immunofluorescence, Staining, Western Blot

Journal: iScience

Article Title: PKA-mediated BMAL1 phosphorylation promotes β 1 -adrenoceptor autoantibody-induced cardiomyocyte death

doi: 10.1016/j.isci.2025.112786

Figure Lengend Snippet: β 1 -AA increases Ser42 phosphorylation of BMAL1 in cardiomyocytes (A and C) Expression of Ser42-phosphorylated BMAL1 (pSer42-BMAL1) in myocardial tissues from control and β 1 -AA groups at different time points. ∗ p < 0.05 vs. control; ∗∗ p < 0.01 vs. control; n = 6. (B and D) pSer42-BMAL1 expression in H9c2 cells in the presence or absence of β 1 -AA at different time points. ∗ p < 0.05 vs. control; n = 5. (E) Expression levels of pSer42-BMAL1 in nuclear and cytosolic fractions of H9c2 cells at CT8 were detected by western blot, with GAPDH (cytoplasm) and histone H3 (nucleus) serving as markers. Data are represented as mean ± SEM.

Article Snippet: The following primary antibodies were used: BMAL1, phospho-BMAL1 (Ser42), Histone H3, phospho-PKA substrate (Cell Signaling Technology, USA); PRKACA (Immunoway, China); Tubulin (Proteintech, China) and GAPDH (Bioss, China).

Techniques: Phospho-proteomics, Expressing, Control, Western Blot