Journal: The Journal of investigative dermatology

Article Title: Impaired trafficking of the desmoplakins in cultured Darier's disease keratinocytes.

doi: 10.1046/j.1523-1747.2003.12557.x

Figure Lengend Snippet: Figure 6. Dp associates with SERCA2 during di¡erentiation. (A) NHK cells grown in low-calcium medium were pulse-labeled with [35S]methionine and cysteine and then chased in a normal-calcium med- ium for 4 h. Cells were lyzed and immunoprecipitated with the indicated antibody. (B) NHK cells were incubated in normal-calcium medium for 4 h to induce cell di¡erentiation. Cells were lyzed and immunoprecipitated with anti-SERCA2 antibody. SERCA2 immunoprecipitates were analyzed by SDS-PAGE and western blot using an anti-Dp antibody (lane 2). Lane 1, Western blot of total cell extract.

Article Snippet: Mouse monoclonal anti-PG antibody was purchased from Cymbus Bioscience (Southampton, UK), goat polyclonal anti-SERCA2 antibody was from Santa Cruz Biotechnology (Insight Biotechnology, Middlesex, UK), and rabbit polyclonal anti-calnexin antibody was obtained from Calbiochem (CN Biosciences, Nottingham, UK).

Techniques: Labeling, Immunoprecipitation, Incubation, SDS Page, Western Blot

Journal: Circulation Research

Article Title: Effects of Atrial Fibrillation on the Human Ventricle

doi: 10.1161/CIRCRESAHA.121.319718

Figure Lengend Snippet: Atrial fibrillation (AF)-simulation in induced pluripotent stem cell cardiomyocytes (iPSC-CMs). Human iPSC-CMs treated either with AF-simulation (arrhythmic pacing: Arr; 60 bpm, 40% beat-to-beat-variability) or rhythmic pacing (control [Ctrl]; 60 bpm) chronically for 7 d. A , Representative recordings of stimulated Ca 2+ transients (epifluorescence microscopy, Fura-2) and ( B ) mean values for Ca 2+ transient amplitude, ( C ) diastolic Ca 2+ levels, ( D ) time to peak 80%, ( E ) relaxation time 80% of human iPSC-CM upon chronic AF-simulation (n=69 cardiomyocytes/6 differentiations/4 donors) or rhythmic pacing (n=71/6/4). F , Original recordings of caffeine-induced Ca 2+ transients (10 mmol/l caffeine, epifluorescence microscopy, Fura-2), ( G ) mean caffeine-transient amplitude indicating the sarcoplasmic reticulum Ca 2+ load and ( H ) SERCA2a activity (K sys -K caff ) of iPSC-CM after chronic AF-simulation (n=13/6/4) vs control (n=12/6/4). I , Representative confocal line scans (Fluo-4) showing diastolic sarcoplasmic reticulum Ca 2+ sparks and ( J ) mean Ca 2+ spark frequency (CaSpF) after chronic AF-simulation (n=68/7/4) vs control (n=67/7/4). K , Original recordings of cytosolic Na + levels (epifluorescence microscopy, SBFI) and ( L ) mean values of cytosolic Na + concentration of human iPSC-CM after chronic AF-simulation (n=110/7/4) compared with control (98/7/4). Data are provided as scatter plot with mean±SD. Each data point is calculated as mean value per differentiation. P were calculated using Student t test ( A–C , E–L ) or Mann-Whitney U test ( D ).

Article Snippet: The expression of RYR2 (ryanodine-receptor type 2), NCX (Na + -Ca 2+ exchanger), SERCA2a, PLB (phospholamban), and CaMKII was studied using specific antibodies anti-RyR2 (mouse monoclonal antibody, dilution 1:1000, Santa Cruz Biotechnology), NCX (mouse monoclonal antibody, dilution 1:1000, Swant), SERCA2a (rabbit polyclonal antibody, dilution 1:1000, Alomone), PLB (mouse monoclonal antibody, dilution 1:1000, Thermo Fisher) and CaMKII-δ (rabbit polyclonal antibody, dilution 1:1000, Thermo Fisher).

Techniques: Epifluorescence Microscopy, Activity Assay, Concentration Assay, MANN-WHITNEY

Journal: Circulation Research

Article Title: Effects of Atrial Fibrillation on the Human Ventricle

doi: 10.1161/CIRCRESAHA.121.319718

Figure Lengend Snippet: Molecular remodeling in the atrial fibrillation (AF) ventricle. Original representative Western Blots of human left ventricular (LV) myocardium from aortic stenosis patients with preserved LV function with sinus rhythm (SR, n=6-7) or AF (n=7) and expression levels (normalized to SR) for ( A ) ryanodine receptor type 2 (RyR2), ( B ) RyR2 phosphorylation at Ser2814 (normalized to total RyR2 expression), ( C ) NCX (Na + /Ca 2+ exchanger), ( D ) SERCA (sarcoplasmic reticulum Ca 2+ ATPase 2a), and ( E ) PLB (phospholamban). Representative Western Blots for ( F ) CaMKII (Ca 2+ /calmodulin-dependent protein kinase IIδc), ( G ) CaMKII phosphorylation at Thr287 (CaMKII-P), and ( H ) CaMKII oxidation at Met281/282 (CaMKII-ox). GAPDH was used as loading control. I , CaMKII activity (CycLex CaMKII activity ELISA kit) and ( J ) H 2 O 2 levels (colorimetric peroxidase assay) in LV myocardium from patients with SR or AF (n=6–7 each). Data are provided as scatter plot with mean±SD. Groups were statistically analysed using Student t test or Mann-Whitney U test (for E and G ).

Article Snippet: The expression of RYR2 (ryanodine-receptor type 2), NCX (Na + -Ca 2+ exchanger), SERCA2a, PLB (phospholamban), and CaMKII was studied using specific antibodies anti-RyR2 (mouse monoclonal antibody, dilution 1:1000, Santa Cruz Biotechnology), NCX (mouse monoclonal antibody, dilution 1:1000, Swant), SERCA2a (rabbit polyclonal antibody, dilution 1:1000, Alomone), PLB (mouse monoclonal antibody, dilution 1:1000, Thermo Fisher) and CaMKII-δ (rabbit polyclonal antibody, dilution 1:1000, Thermo Fisher).

Techniques: Western Blot, Expressing, Activity Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY