Journal: Biomedicines

Article Title: Prior Aerobic Exercise Training Fails to Confer Cardioprotection Under Varying Exercise Volumes in Early Post-Infarction Cardiac Remodeling in Female Rats

doi: 10.3390/biomedicines13092221

Figure Lengend Snippet: Calcium-handling proteins. ( A ) PLB, total phospholamban; ( B ) PLB(Thr17/Ser16), phospholamban phosphorylated at serine 16 and threonine 17; ( C ) NCX1, sodium/calcium exchanger 1; ( D ) SERCA2a, sarco/endoplasmic reticulum Ca 2+ -ATPase; ( E ) LTCC, L-type Ca v 1.2 voltage-gated calcium channel; ( F ) RyR, ryanodine receptor. GAPDH: glyceraldehyde 3-phosphate dehydrogenase constitutive protein. Boxplots (black squares with whiskers) with individual data points (small circles of different colors) and means (red circles) or 20% trimmed means (red squares). The p -value displayed above the boxplots refers to the ANOVA. Brackets were added to the graphs to represent significant differences in pairwise comparisons. ( B – D , F ) charts: Fisher’s one-way ANOVA and pairwise comparisons via Student’s t test. ( A , E ) charts: Welch’s ANOVA with trimmed means and pairwise comparisons via Yuen’s trimmed test. The GAPDH-LTCC and GAPDH-RyR are equivalent because the same membrane was used to detect LTCC and RyR through membrane stripping for Western blotting.

Article Snippet: The transferred proteins were then incubated at ~4 °C (overnight) with the following primary rabbit antibodies: anti-L-type high-voltage calcium channel Ca v 1.2 (LTCC) (1:10,000 ACC-003, Alomone, Jerusalem, Israel), anti-sarco/endoplasmic reticulum Ca 2+ -ATPase (SERCA2a) (1:1000 A010-20, Badrilla, Leeds, UK), anti-phospholamban phosphorylated at threonine 17 and serine 16 (PLB(Thr17/Ser16)) (1:5000 ab62170, Abcam, Cambridge, MA, USA), anti-sodium/calcium exchanger 1 (NCX1) (1:5000 ab177952, Abcam, Cambridge, MA, USA), anti-4-hydroxynonenal (4-HNE) (1:4000 ab46545, Abcam, Cambridge, MA, USA), and anti-GAPDH (1:20,000 D16H11, Cell Signaling Technology, Danvers, MA, USA), as well as the following primary mouse antibodies: anti-ryanodine receptor (RyR) (1:2000 ab2868, Abcam, Cambridge, MA, USA), and anti-total phospholamban.

Techniques: Membrane, Stripping Membranes, Western Blot