Journal: Cell Research
Article Title: A GABAergic system in atrioventricular node pacemaker cells controls electrical conduction between the atria and ventricles
doi: 10.1038/s41422-024-00980-x
Figure Lengend Snippet: a Illustrator and workflow for the single-cell gene expression analysis of AVNPCs from Hcn4 CreERT2(+) ; Rosa26 TomRed+ mice. b Heatmap showing GABAergic system gene expression in single AVNPCs from Hcn4 CreERT2(+) ; Rosa26 TomRed+ mice. Rows indicate the AVNPC samples, and columns indicate the cycle threshold (Ct) values of GABAergic system genes. Ct values were scaled and the relative gene expression was demonstrated using color scales. A score of –1 (red) indicates a high expression level, and a score of 1 (blue) indicates a low level of expression. The data were obtained from 52 AVNPCs from 5 Hcn4 CreERT2(+) ; Rosa26 TomRed+ mice. c Immunofluorescence staining showing the expression and localization of GABA metabolic enzymes (GAD2, GABA-T, and SSADH), GABA A receptors (GABRA3, GABRB2, and GABRG2), GABA transporters (vGAT and GAT-1) in single mouse AVNPCs. Scale bar, 10 μm.
Article Snippet: The following primary antibodies were used in immunofluorescence experiments: anti-GABA antibody (5A9) (Abcam, ab86186, 1:100 for cells); anti-GABRA3 antibody (Alomone, AGA-003, 1:200 for cells, 1:50 for heart slices); anti-GABRB2 antibody (Invitrogen, PA5-64331, 1:100 for cells, 1:20 for heart slices); anti-GABRG2 antibody (Alomone, AGA-005, 1:100 for cells, 1:50 for heart slices); anti-GAT-1 antibody (Synaptic Systems, 274102, 1:200 for cells, 1:50 for heart slices); anti-vGAT antibody (Alomone, AGT-005, 1:100 for cells, 1:50 for heart slices); anti-GAD2 antibody (Santa Cruz Biotechnology, sc-377145, 1:100 for cells, 1:50 for heart slices); anti-GABA-T antibody (Abcam, ab216465, 1:100 for cells, 1:50 for heart slices); anti-SSADH antibody (Santa Cruz Biotechnology, sc-390754, 1:100 for cells, 1:50 for heart slices); anti-CX43 antibody (Cell Signaling Technology, 3512, 1:50 for heart slices); anti-HCN4 antibody produced in mouse (Sigma, SAB5200035, 1:100 for cells, 1:50 for heart slices); anti-HCN4 antibody produced in rat (Sigma, MABN1871, 1:100 for cells, 1:50 for heart slices); and anti-CAST antibody (Invitrogen, PA5-87352, 1:100 for cells, 1:50 for heart slices).
Techniques: Expressing, Immunofluorescence, Staining