Fig 2: ATP hydrolysis activity corresponds to p53 peaks in FPLC gel filtration profile.The FPLC profile of p53 shows two peaks for UV absorbance at 280(in Red) corresponding to higher oligomeric form (peak 1) and monomeric form (peak 2). The peak fractions 47 and 63 were loaded on SDS-PAGE gel and both show the same size band corresponding to p53-GST (Inset). Equal volume of various fractions collected from FPLC elution was taken to perform NADH dependent ATP hydrolysis assay. The rates of ATP hydrolysis (Absorbance decline at 340 nm per min) are plotted on the secondary Y axis as bar graph (in Blue) to compare the protein concentration and ATPase activity for each fraction.

Fig 3: Targeting of HDAC4 and FOXP1 by miR-22 and constitutive acetylation of p53 may contribute to the overexpression of p21 and/or p27.a Diagram of the “seed” sequence for miR-22 targeting of HDAC4 and FOXP1. b Whole cell lysates prepared from S05 and 182R-6 cells were subjected to western blot analysis using antibodies to either HDAC4 or FOXP1. GAPDH served as a loading control. c 182R-6 cells transfected with either 40 nM miR-22 mimic or AllStars negative control siRNA or 50 nM miR-22 inhibitor or negative control were incubated for either 72 or 96 h. Whole cellular lysates were prepared and subjected to western blot analysis using the indicated antibodies. GAPDH served as a loading control. d 182R-6 cells transfected with either 40 nM miR-22 mimic or AllStars negative control siRNA or 50 nM miR-22 inhibitor or negative control were incubated for either 72 or 96 h. Whole cell lysates were prepared, and the immunoprecipitation and western blot analysis were performed as described in the “Methods” section

Fig 4: HER2/NF-κB (p65)/miR-22/HDAC4/p21 and HER2/NF-κB (p65)/miR-22/Ac-p53/p21 signaling circuits contribute to the dual role of miR-22 in resensitizing fulvestrant-resistant breast cancer cells to fulvestrant by transactivation of p21.a The HER2/p65 dephosphorylation (ser536) axis negatively modulates miR-22 transcription. b Gain-of-function of miR-22 represses the expression of HDAC4 and FOXP1, which leads to attenuation of the transcription repressor complex63, resulting in a transcriptional activation of p21. c Loss-of-function of miR-22 induces acetylation of p53 at Lys382, resulting in a transactivation of p21

Fig 5: Model of iPSC generation from human keratinocytesNotch inhibition allows the production of safer oncogene-free iPSCs by suppressing p21 in a p53-independent manner.