Detection of CDKN3 protein by immunofluorescence and Western blot in cell lines derived from CC transfected with specific siRNAs against <i>CDKN3</i> or scrambled siRNAs.

<p>Cell lines derived from cervical cancer (CC) positive for human papilloma virus (HPV) 16 (CaSki, SiHa) and HPV18 (HeLa) were transfected with specific cyclin-dependent kinase inhibitor 3 <i>(CDKN3)</i> or scrambled siRNAs. Cells were harvested at 96 h after transfection and stained for CDKN3 protein. (A) Immunofluorescence staining for CDKN3 protein in SiHa, CaSki and HeLa cell lines using an anti-CDKN3 primary antibody and FITC-conjugated secondary antibody. Images were photographed at 60× magnification using a fluorescence microscope Olympus FV 1000. (B) Quantification of fluorescence intensity of anti-CDKN3 antibody-stained cells. The green fluorescence intensity was quantified using ImageJ software. Values represent the mean ± S.D. of 140 fields measured in each experiment. The statistical significance between the differences was calculated using the t test. (C) Expression of CDKN3 protein examined using western blot in SiHa, CaSki, and HeLa cell lines transfected with random siRNAs (-) and with specific <i>CDKN3</i> siRNAs (+) with actin as internal control.