erk5 upstate  (Cell Signaling Technology Inc)

Journal: Journal of Molecular and Cellular Cardiology

Article Title: Targeted deletion of ERK2 in cardiomyocytes attenuates hypertrophic response but provokes pathological stress induced cardiac dysfunction

doi: 10.1016/j.yjmcc.2014.03.002

Figure Lengend Snippet: Characterisation of cardiomyocyte-specific deletion of ERK2. (A) Quantitative real-time PCR analysis of mRNA levels of ERK2 in the left ventricle (LV), skeletal muscle (SM), brain, and liver. A 73% decrease in mRNA level was shown only in LV of ERK2 cko mice in comparison to ERK2 f/f mice. Data were derived from 3 independent experiments performed in triplicate and normalized to GAPDH content ( n = 3). (B) Immunoblot analysis confirmed specificity of ERK2 deletion in LV. ERK2 protein expression was comparable in various tissues. Tubulin served as protein loading control. The ratio of ERK2 expression to tubulin is shown in the bar graph. (C) Western blot analyses demonstrated unchanged activation and expression levels of ERK1 and MEK1/2. Expression of ERK3, ERK4, ERK5, p38 and JNK in ventricular extracts was the same in all genotypes. Tubulin served as protein loading control. (D) ERK1/2 kinase activity was examined on the tissue subjected to 1 week of TAC, demonstrating ERK1 activation did not compensate the loss of ERK2 in the heart ( n = 5). n.s.: no significant difference. Data presented as mean ± SEM.

Article Snippet: Protein extracts (30 μg) were subjected to Western blot analyses with the primary antibodies against ERK1 (Santa Cruz), ERK2 (Santa Cruz), ERK5 (Upstate), Phospho-ERK5 (Invitrogen), p38 (Santa Cruz), Phospho-ERK1/2, MEK1/2, Phospho-MEK1/2, Phospho-p38, JNK1/2, Phospho-JNK1/2, PKB, Phospho-PKB, active caspase 3 (Cell signaling) or α-Tubulin (Sigma-Aldrich).

Techniques: Real-time Polymerase Chain Reaction, Derivative Assay, Western Blot, Expressing, Activation Assay, Activity Assay

Journal: Journal of Molecular and Cellular Cardiology

Article Title: Targeted deletion of ERK2 in cardiomyocytes attenuates hypertrophic response but provokes pathological stress induced cardiac dysfunction

doi: 10.1016/j.yjmcc.2014.03.002

Figure Lengend Snippet: Analysis of hypertrophic regulators in ERK2 f/f and ERK2 cko ventricles. Protein extracts from ERK2 f/f and ERK2 cko ventricles after 1 week of sham or TAC operation were subjected to immunoblot analyses for total ERK2, ERK1, MEK1/2, PKB, p38, JNK and ERK5 expression as well as their phosphorylation levels using specific antibodies.

Article Snippet: Protein extracts (30 μg) were subjected to Western blot analyses with the primary antibodies against ERK1 (Santa Cruz), ERK2 (Santa Cruz), ERK5 (Upstate), Phospho-ERK5 (Invitrogen), p38 (Santa Cruz), Phospho-ERK1/2, MEK1/2, Phospho-MEK1/2, Phospho-p38, JNK1/2, Phospho-JNK1/2, PKB, Phospho-PKB, active caspase 3 (Cell signaling) or α-Tubulin (Sigma-Aldrich).

Techniques: Western Blot, Expressing