HIPPI interacts with P53 in the nucleus and alters the expression of Procaspase1 in cell. A. Co-immunoprecipitation of HIPPI with P53 from whole cell extract of Neuro2A cells. IP was carried out with anti P53 antibody and western blot was done with anti HIPPI antibody (Upper panel). The same blot was stripped and reprobed with anti P53 antibody as IP control (lower panel). Lane 'In': input of immunoprecipitation, '+Ab': cell extract treated with anti P53 antibody, '-Ab': cell extract treated with IgG only. B. Co-immunoprecipitation of HIPPI with P53 from cytoplasm and nuclear compartment of Neuro2A cells. Cytoplasmic ('CE') and nuclear ('NE') fractions were separately immunoprecipitated with anti P53 antibody and detected with anti HIPPI antibody (Upper panel). The lane markings carry their usual meanings as described in A. C. Interaction of HIPPI with P53 in cytoplasm and nuclear compartment of N2AHip1Si cells. D. Western blot analysis for the detection of Procaspase1 expression in parental HeLa cells ('HeLa'), HeLa cells transfected with GFP-Hippi ('Hippi'), p53 knocked down HeLa cells exogenously transfected with GFP-Hippi ('P53SiHi'), HeLa cells transfected with full length p53 ('P53') and HeLa cells co-transfected with full length p53 and GFP-Hippi ('P53+Hippi'). Beta actin was used as loading control. E. Bar diagram represents mean (n = 3) relative expression of Procaspase1 in different cells described in D above. IOD value of the Procaspase1 specific band was normalized with that of beta actin. Normalized IOD of 'HeLa' was considered to be 1.