Fig. 7.

Role of p38/MAPK activation in MAP4 phosphorylation-induced cardiac hypertrophy, apoptosis and fibrosis. (a) Detection of p-M and p-P38 with or without SB203580 (SB) (5 μM) in CMs. n = 5. (b) Protein markers of hypertrophy and p-P38 were detected using WB. n = 5. (c) Cross sectional areas of CMs were analyzed using cTnI and FITC-WGA co-staining. n = 5. (d and e) CMs apoptosis was detected by cTnI and fluorescein-TUNEL co-staining. Bar, 10 μm. n = 5. (f) Protein levels of caspase-3 and t-caspase-3 were analyzed using WB. n = 5. (g) Detection of Cyt-c release from mitochondria. n = 5. (h) CMs mitochondria observed by TEM (black square indicated representative mitochondria). Bar, 1 μm. (i) Markers of ECM were examined using WB. n = 5. (j) Poly/free tubulin was analyzed using WB. n = 5. (k) Representative confocal immunofluorescence images showing the MTs organization of the CMs. Bar, 10 μm. The inserts showed high magnification images of the peripheral MT network. (l) Detection of p-M translocation to mitochondria with or without SB. n = 5. (m and n) CMs apoptosis was detected by cTnI and fluorescein-TUNEL co-staining after the cells were transiently transfected with MAP4(Ala), MKK6(Glu) or both. Bar, 10 μm. n = 5. (o) Representative confocal immunofluorescence images showing the MTs organization of the CMs after the cells were transiently transfected with MAP4(Ala), MKK6(Glu) or both. Bar, 10 μm. The inserts showed high magnification images of the peripheral MT network. (p) CMs mitochondria observed by TEM (black square indicated representative mitochondria) after the cells were transiently transfected with MAP4(Ala), MKK6(Glu) or both. Bar, 1 μm. Data were shown as mean ± SEM. The experiment was conducted 5 times. *P < 0.05, **P < 0.01, ***P < 0.001. P values were derived from one-way ANOVA with Bonferroni's post-test.