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: PLoS ONE

Article Title: Low Density Lipoproteins Promote Unstable Calcium Handling Accompanied by Reduced SERCA2 and Connexin-40 Expression in Cardiomyocytes

doi: 10.1371/journal.pone.0058128

Figure Lengend Snippet: Real-time PCR analysis showing SERCA2 (A), RyR2 (B), IP3-RI (C) and IP3-RIII mRNA expression levels in HL-1 cardiomyocytes exposed to increasing doses of LDL. Data were processed with a specially designed software programme based on Ct value of each sample and normalized to ARBP mRNA. Values significantly different from the level with 0 µg/ml LDL are indicated with asterisks.

Article Snippet: Blots were incubated with monoclonal antibodies against SERCA2 (Novus Biologicals NB100–237), Cx40 (Invitrogen, 36–4900) and Cx43 (Sigma-Aldrich C6219).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Software

Journal: PLoS ONE

Article Title: Low Density Lipoproteins Promote Unstable Calcium Handling Accompanied by Reduced SERCA2 and Connexin-40 Expression in Cardiomyocytes

doi: 10.1371/journal.pone.0058128

Figure Lengend Snippet: Representative Western blot analysis showing SERCA2 bands in HL-1 cells exposed to increasing LDL concentrations (0, 50, 100 and 200 µg/mL) ( A ) or to similar dose (500 µg/ml) of LDL or HDL ( B ) for 24 hours. The bar graph below shows SERCA2 band quantification. Unchanged levels of β-tubulin are shown as loading control. Results are mean±SEM of three independent experiments performed in duplicate. *P<0.05 versus. HL-1 cells incubated in absence of LDL. C Calcium images acquired at rest before (CON) and during a rapid caffeine application. D Representative calcium transients recorded in cultures incubated without lipoproteins (CON), with 500 µg/ml LDL or with 500 µg/ml HDL. The lower panel shows the corresponding time integrals of the calcium transients. E summary of the effect of LDL and HDL on the time integral of the caffeine induced calcium transient. *P<0.05 vs. cells incubated with LDL

Article Snippet: Blots were incubated with monoclonal antibodies against SERCA2 (Novus Biologicals NB100–237), Cx40 (Invitrogen, 36–4900) and Cx43 (Sigma-Aldrich C6219).

Techniques: Western Blot, Control, Incubation

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Calotropin attenuates ischemic heart failure after myocardial infarction by modulating SIRT1/FOXD3/SERCA2a pathway.

doi: 10.1016/j.biopha.2024.117384

Figure Lengend Snippet: Fig. 4. SIRT1 inhibitor EX527 blocked the cardioprotective effects of CAL against HF after MI. (A) Representative M-mode images of rats from each group. (B to E) The effect of CAL on the parameters of cardiac function in the presence of EX527. n = 6; All data are presented as mean ± SEM and analyzed using One-way ANOVA follow by Tukey post-hoc tests; # P<0.05, ### P<0.001, vs Sham group; ** P<0.01, *** P<0.001, vs HF group or HF+CAL+EX527 group. (F) Representative hematoxylin–eosin(H&E) staining images of cardiac tissue. (G and H) Representative Masson staining images of cardiac tissue and the analysis of fibrotic areas. n = 6; The data were expressed as mean ± SEM and analyzed using One-way ANOVA follow by Tukey post-hoc tests; ### P<0.001, vs Sham group; *** P<0.001, vs HF group or HF+CAL+EX527 group.

Article Snippet: Then they were incubated with primary antibodies against SERCA2a (1:1000, A11692, ABclonal), SIRT1 (1:1000, A0230, ABclonal), forkhead Box D3 (FOXD3) (1:1000, 5G9B10, Santa Cruz), GAPDH (1:10000, AC010, ABclonal), and β-Actin (1:100000, A026, ABclonal).

Techniques: Staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Calotropin attenuates ischemic heart failure after myocardial infarction by modulating SIRT1/FOXD3/SERCA2a pathway.

doi: 10.1016/j.biopha.2024.117384

Figure Lengend Snippet: Fig. 5. CAL preserved SERCA2a expression via SIRT1 enhancement. (A to C) The effect of CAL on the protein expression of SIRT1 and SERCA2a in HF rat car diomyocytes with EX527 treatment or without. n = 5; The data were expressed as mean ± SEM and analyzed using One-way ANOVA follow by Tukey post-hoc tests; # P<0.05, ## P<0.01, vs Sham group; * P<0.05, ** P<0.01, vs HF group or vs HF+CAL+EX527 group. (D) The effect of CAL on the mRNA expression of SERCA2a in HF rat cardiomyocytes with EX527 treatment or without. n = 5; The data were expressed as mean ± SEM and analyzed using One-way ANOVA follow by Tukey post- hoc tests; ### P<0.001, vs Sham group; *** P<0.001, vs HF group or vs HF+CAL+EX527 group. (E to G) The effect of CAL on the protein expression of SIRT1 and SERCA2a in OGD/R-induced H9c2 cells with EX527 treatment or without. n = 3; The data were expressed as mean ± SD and analyzed using One-way ANOVA follow by Tukey post-hoc tests; # P<0.05, ## P<0.01, vs Sham group; * P<0.05, ** P<0.01, vs HF group or vs HF+CAL+EX527 group. (H) The effect of CAL on the mRNA expression of SERCA2a in OGD/R-induced H9c2 cells with EX527 treatment or without. n = 3; (I-K) The effect of SIRT1 silence on the increased expression of SERCA2a induced by CAL. The data were expressed as mean ± SEM and analyzed using One-way ANOVA follow by Tukey post-hoc tests; ### P<0.001, vs Sham group; *** P<0.001, vs HF group or vs HF+CAL+EX527 group.

Article Snippet: Then they were incubated with primary antibodies against SERCA2a (1:1000, A11692, ABclonal), SIRT1 (1:1000, A0230, ABclonal), forkhead Box D3 (FOXD3) (1:1000, 5G9B10, Santa Cruz), GAPDH (1:10000, AC010, ABclonal), and β-Actin (1:100000, A026, ABclonal).

Techniques: Expressing

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Calotropin attenuates ischemic heart failure after myocardial infarction by modulating SIRT1/FOXD3/SERCA2a pathway.

doi: 10.1016/j.biopha.2024.117384

Figure Lengend Snippet: Fig. 7. CAL promoted SERCA2a transcription via SIRT1-dependent deacetylation of FOXD3 in HF post-MI. (A) The effect of CAL on FOXD3 expression in the presence of EX527 in LAD-induced HF rat heart tissue. (B) The effect of CAL on FOXD3 expression in the presence of EX527 in OGD/R-induced H9c2 cells. (C-F) The effect of CAL on the FOXD3 location. ##P<0.01, ### P<0.001, vs control group; ** P<0.01, vs OGD/R group. (G) Immunofluorescence images for FOXD3. (H)The effect of CAL on FOXD3 acetylation in the presence of EX527 in LAD-induced HF rat heart tissue. (I) The effect of CAL on FOXD3 acetylation in the presence of EX527 in OGD/ R-induced H9c2 cells.

Article Snippet: Then they were incubated with primary antibodies against SERCA2a (1:1000, A11692, ABclonal), SIRT1 (1:1000, A0230, ABclonal), forkhead Box D3 (FOXD3) (1:1000, 5G9B10, Santa Cruz), GAPDH (1:10000, AC010, ABclonal), and β-Actin (1:100000, A026, ABclonal).

Techniques: Expressing, Control, Immunofluorescence

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Calotropin attenuates ischemic heart failure after myocardial infarction by modulating SIRT1/FOXD3/SERCA2a pathway.

doi: 10.1016/j.biopha.2024.117384

Figure Lengend Snippet: Fig. 8. Calotropin improved cardiac function and prevented adverse ventric ular remodeling in ischemic heart failure after MI via the SIRT1/FOXD3/SER CA2a pathway.

Article Snippet: Then they were incubated with primary antibodies against SERCA2a (1:1000, A11692, ABclonal), SIRT1 (1:1000, A0230, ABclonal), forkhead Box D3 (FOXD3) (1:1000, 5G9B10, Santa Cruz), GAPDH (1:10000, AC010, ABclonal), and β-Actin (1:100000, A026, ABclonal).

Techniques: