Research Article
P16INK4a Regulates ROS-Related Autophagy and CDK4/6-Mediated Proliferation: A New Target of Myocardial Regeneration Therapy
Figure 6
P16INK4a mediates abnormal accumulation of ROS and autophagy. (a) The mean fluorescence intensity of DCF in cardiomyocytes transfected with Ad5:cTnT-INK4a (INK4a) or Ad5:cTnT-CON (NC) was determined by immunofluorescence in vitro (). Scale bar: 20 μm. (b) The mean fluorescence intensity of DCF in cardiomyocytes transfected with INK4a or NC was determined by immunofluorescence at 6 dpr (). Scale bar: 20 μm, 50 μm. (c) DNA damage is quantified by immunofluorescence for the staining of γH2X in cardiomyocytes at 6 dpr (). Scale bar: 20 μm, 50 μm. (d) The expression of p16INK4a, Beclin1, ATG5, and LC3B in the ventricular muscle after specific overexpression of p16INK4a in 6dpr. (e) The number of autophagosomes was statistically analysed by immunofluorescence staining in INK4a or NC group (). Scale bar: 20 μm. (f) The effect of p16INK4a overexpression on autophagosomes of mice cardiomyocytes during 28 dpr was detected by transmission electron microscopy () (data are presented as , , , ).
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