Journal: Cell

Article Title: Multidimensional tracking of GPCR signaling via peroxidase-catalyzed proximity labeling

doi: 10.1016/j.cell.2017.03.028

Figure Lengend Snippet: (A) Internalization kinetics of β2AR (class A GPCR) are different from those of AT1R (class B GPCR). β-arrestin is recruited with the peak at 180 s, but clathrin recruitment is delayed to 600 s. Also, Rab5 enrichment peak is at 600 s, indicating arrestin-receptor complex formation and endosomal entry do not occur simultaneously. Late endosomal marker Rab7 and lysosomal marker LAMP1 enrichment peaks appear afterward.

Article Snippet: Each well was transfected with 20 ng each of a pcDNA5 encoding β2AR-APEX/β2AR wild-type and CRE-SEAP reporter plasmid (BD Biosciences).

Techniques: Marker

Journal: Biochemical and biophysical research communications

Article Title: Myofibroblast β2 adrenergic signaling amplifies cardiac hypertrophy in mice

doi: 10.1016/j.bbrc.2019.01.070

Figure Lengend Snippet: (A) Representative images of hematoxylin-eosin staining of whole hearts from wildtype control or β2ARKO mice 2 weeks after isoproterenol (ISO) stimulation or sham operation. The bar indicates 1 mm. (B) Gravimetric analysis. Heart weight (HW) normalized to body weight (BW) ratios are shown. (C) Lung weight normalized to BW (LW/BW) ratios. (D) Fractional shortening (%) obtained 2 weeks after sham operation or ISO-pump implantation by echocardiographic analysis. (E) Left ventricular dimension at end-diastole (LVEDd) obtained 2 weeks after sham operation or ISO-pump implantation by echocardiographic analysis. (F) Representative image of WGA-FITC conjugate staining of heart tissue. The bar indicates 50 μm. (G) Cross-sectional area measurements in WGA-FITC conjugate-stained hearts. (H, I, J) Expression levels of cardiac hypertrophic marker genes, Nppa (H), Nppb (I), and Acta1 (J), in ventricles from mouse cohorts determined by real-time RT-PCR. Values were normalized to that of GAPDH and are represented as fold increases relative to that in the wildtype sham group. Values are shown as mean ± SD, and the number displayed on each column indicates the number of samples. † P<0.05 vs. all other groups, *P<0.05 between two indicated groups by one-way ANOVA followed by Tukey–Kramer test.

Article Snippet: Genetically engineered mice β2AR null mice (β2ARKO) were obtained from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Staining, Control, Expressing, Marker, Quantitative RT-PCR

Journal: Biochemical and biophysical research communications

Article Title: Myofibroblast β2 adrenergic signaling amplifies cardiac hypertrophy in mice

doi: 10.1016/j.bbrc.2019.01.070

Figure Lengend Snippet: (A) Representative images of M-T staining of heart tissue obtained from wildtype or β2ARKO mice at 2 weeks after sham-operation or ISO-stimulation. The bar indicates 100 μm. (B) Quantification of fibrotic area assessed by M-T staining. (C, D) Expression levels of cardiac fibrotic marker gene, Col1a2 (C) and Col3a1 (D) in ventricles determined by real-time RT-PCR. Values were normalized to that of GAPDH and are represented as fold increases relative to that in the wildtype sham group. (E) Representative images of cell migration as assessed by scratch assay. Images of scratched confluent cardiac fibroblasts from wildtype control (upper panels) or β2ARKO (lower panels) mice were taken at indicated time points. Arrows indicate the width of cell-free areas. The bar indicates 200 μm. (F) Quantitation of migration distance of cardiac fibroblasts at 6 h and 12 h. Values are mean ± SD, and the number displayed on each column indicates the number of samples. †P<0.05 vs. all other groups, *P<0.05 between two indicated groups by one-way ANOVA followed by Tukey-Kramer test.

Article Snippet: Genetically engineered mice β2AR null mice (β2ARKO) were obtained from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Staining, Expressing, Marker, Quantitative RT-PCR, Migration, Wound Healing Assay, Control, Quantitation Assay

Journal: Biochemical and biophysical research communications

Article Title: Myofibroblast β2 adrenergic signaling amplifies cardiac hypertrophy in mice

doi: 10.1016/j.bbrc.2019.01.070

Figure Lengend Snippet: (A) Representative image of βAR RT-PCR products obtained from adult mouse cardiomyocytes or fibroblasts. (B) Representative images of neonatal rat cardiomyocytes (NRCMs) with actinin staining 24 h after stimulation with ISO (0.5 μM or 10 μM) or vehicle. (C) Cell surface area measurements of NRCMs 24 h after stimulation with low or high dose of ISO (0.5 μM or 10 μM) normalized to those of NRCMs treated with vehicle. Values are mean ± SD of three independent experiments. † P<0.05 vs. other groups by Student’s t-test. (D) Representative images of actinin staining of NRCMs stimulated with media conditioned by cardiac fibroblasts from wildtype or β2ARKO mice with or without ISO treatment. (E) Cell surface area measurements of NRCMs stimulated with media conditioned by cardiac fibroblasts from wildtype or β2ARKO mice with or without ISO treatment. The values were normalized to those of NRCMs treated with media conditioned by wildtype without ISO treatment. Values are mean ± SD of four independent experiments. P<0.05 between two indicated groups by one-way ANOVA followed by Tukey-Kramer test. The bars indicate 50 μm.

Article Snippet: Genetically engineered mice β2AR null mice (β2ARKO) were obtained from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Reverse Transcription Polymerase Chain Reaction, Staining