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. 2012 Dec 21;7(12):e51427. doi: 10.1371/journal.pone.0051427

Figure 3. Synthesis of cyclin D, CDK4, p16INK4 and p53 in HCEnC-21 and HCEnC-21T.

Figure 3

(A) Analysis of p53 functionality. Oxidative stress was induced in confluent monolayers of HCEnC-21 and HCEnC-21T cells using 50 µM tert-Butyl-hydroperoxide for 1 hr. Cell extracts were then separated by SDS-PAGE and immunoblotted with antibodies against p53, phospho-p53 (serine-15), and β-actin. P53 was detected in extracts of both HCEnC-21 and HCEnC-21T and levels of phospho-p53 (activated p53) were increased upon induction of oxidative stress. E: earlier passages <25. L: later passages >45. (B) Synthesis of G1 phase regulatory proteins. Cell extracts were run on SDS-polyacrylamide gels and immunoblotted with antibodies against cyclin D, CDK4, p16INK4 and β-actin. No changes in p16INK4 protein levels were detected. Cyclin D and CDK4 levels were increased in both, HCEnC-21 and HCEnC-21T compared to 21M.