Research Article
P16INK4a Regulates ROS-Related Autophagy and CDK4/6-Mediated Proliferation: A New Target of Myocardial Regeneration Therapy
Figure 7
Alleviation of ROS by NAC relieves p16INK4a-mediated CDK4/6 regulation of NMCM proliferation. (a) The mean fluorescence intensity of DCF in cardiomyocytes treated with Ad5:cTnT-INK4a (INK4a) or INK4a + NAC was determined by immunofluorescence (). Scale bar: 20 μm. (b) The number of autophagosomes was statistically analysed by immunofluorescence staining in INK4a or INK4a + NAC group (). Scale bar: 20 μm. (c) The expression of p16INK4a, CDK4/6, and CyclinD1 in cardiomyocytes treated with INK4a or INK4a + NAC was detected by Western blot. (d) The expression of Beclin1, ATG5, and LC3B in cardiomyocytes treated with INK4a, INK4a + NAC, or INK4a + NAC + RPM was detected by Western blot. (e) The expression of CDK4 and CDK6 in cardiomyocytes treated with NC, Ad5:cTnT-INK4ai (INK4ai), or INK4ai + PAL was detected by Western blot. (f, g) Cardiomyocyte proliferation is quantified by immunofluorescence for Ki67 and pH 3 in cardiomyocytes transfected with NC/INK4ai/INK4ai + PAL (). Scale bar: 50 μm (data are presented as , , , ).
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