Journal: JACC: Basic to Translational Science

Article Title: Acute Glycogen Synthase Kinase-3 Inhibition Modulates Human Cardiac Conduction

doi: 10.1016/j.jacbts.2022.04.007

Figure Lengend Snippet: Acute SB2 Treatment Increases β -Catenin Protein in Human LV Slices (A) Western blot of β-catenin and GAPDH levels with quantification based on band density relative to GAPDH shows an increase in β-catenin (31% average increase) in SB2-treated slices. (B) Both chromatin-bound β-catenin and nonphosphorylated β-catenin levels increased with respect to histone H3 in slices cultured with SB2. In contrast to other subcellular fractions, chromatin-bound β-catenin shows multiple bands of lower molecular weights in addition to the primary band at 92 kDa. (C) ABC (green) is present in human myocyte nuclei, marked by PCM1 (red) and DAPI (blue) , in baseline, 3-hour control, and 3-hour SB2 conditions. White arrowheads indicate β-catenin–positive cardiomyocyte nuclei, whereas white asterisks mark β-catenin–negative cardiomyocyte nuclei. Scale = 50 μm. Statistics were performed using repeated-measures analysis of variance followed by Tukey’s post hoc test for multiple pairwise comparisons. ∗ P < 0.05. ABC = active β-catenin; DAPI = 4′,6-diamidino-2-phenylindole; non-P = nonphosphorylated; other abbreviations as in Figure 2 .

Article Snippet: Immunohistochemistry was performed on optimal cutting temperature compound–embedded frozen sections (Sakura 4583) with a combination of sarcomeric α-actinin antibody (1:100, Abcam ab9465), Na V 1.5 antibody (1:100, Cell Signaling Technology 14421), PCM1 antibody (1:1,000, Sigma HPA023370), active β-catenin (ABC) antibody (1:50, Developmental Studies Hybridoma Bank, University of Iowa PY489-B-catenin), pan β-catenin antibody (1:100, BD 610153), and Cx43 antibody (1:50, Thermo Fisher Scientific 71-0700).

Techniques: Western Blot, Cell Culture

Journal: JACC: Basic to Translational Science

Article Title: Acute Glycogen Synthase Kinase-3 Inhibition Modulates Human Cardiac Conduction

doi: 10.1016/j.jacbts.2022.04.007

Figure Lengend Snippet: Myocyte-Specific β -Catenin GOF and Acute GSK-3 Inhibition by SB2 Reduces Conduction Velocity in Murine Hearts (A) Representative activation maps showing slowed electrical activation in β-catenin GOF hearts (right) compared to noninduced controls (left) . (B) Activation map showing ventricular tachycardia induced from electrical stimulation of β-catenin GOF ventricles. The white arrows show irregular propagation of 2 different wave fronts, which results in a functional block. Five of 6 β-catenin GOF and 0 of 6 control hearts had spontaneous or stimulation-induced ventricular tachycardia. (C) Conduction velocities of RVs and LVs are significantly reduced in β-catenin GOF hearts compared to controls. (D) Western blot shows expression of stabilized β-catenin (lower-molecule-weight band associated with loss of exon 3) in β-catenin GOF mice and decreased Na V 1.5 protein in β-catenin GOF compared to control hearts. (E) Transcription of Wnt/β-catenin target gene Axin2 is increased, and Scn5a is decreased, in β-catenin GOF compared to control hearts. Statistics were performed using Student’s t-test : ∗ P < 0.05; ∗∗ P < 0.01. β-cat = β-catenin; GOF = gain of function; RV = right ventricle; other abbreviations as in Figure 2 .

Article Snippet: Immunohistochemistry was performed on optimal cutting temperature compound–embedded frozen sections (Sakura 4583) with a combination of sarcomeric α-actinin antibody (1:100, Abcam ab9465), Na V 1.5 antibody (1:100, Cell Signaling Technology 14421), PCM1 antibody (1:1,000, Sigma HPA023370), active β-catenin (ABC) antibody (1:50, Developmental Studies Hybridoma Bank, University of Iowa PY489-B-catenin), pan β-catenin antibody (1:100, BD 610153), and Cx43 antibody (1:50, Thermo Fisher Scientific 71-0700).

Techniques: Inhibition, Activation Assay, Functional Assay, Blocking Assay, Western Blot, Expressing

Journal: Frontiers in Immunology

Article Title: A 31-plex panel for high-dimensional single-cell analysis of murine preclinical models of solid tumors by imaging mass cytometry

doi: 10.3389/fimmu.2022.1011617

Figure Lengend Snippet: Antibody Panel.

Article Snippet: β-catenin , D13A1 , Fluidigm , 165 Ho , 1/400 , EMT.

Techniques: Immunopeptidomics