Fig. 3

Stable shRNA knockdown of Oct4A in HEY ovarian cancer cell line. a The efficiency of Oct4A knockdown was evaluated by quantitative real-time PCR analysis. Relative quantification of Oct4A expression was standarized to 18S housekeeping gene and normalised to vector control cells. Data is from three independent samples assessed in triplicate. b The expression and localization of Oct4A in HEY cells following stable shRNA knockdown of Oct4A was also evaluated using immunofluorescence staining. Cells were immunostained using mouse monoclonal Oct4A-specific antibody and visualized using the secondary Alexa 488 fluorescent labelled antibody (green). Nuclear staining was visualized using DAPI staining (blue). Images are representative of three independent experiments. Magnification is set at 200x. Scale bar is set at 50 μM. c Mean fluorescence intensity of Oct4A staining is presented as the mean ± SEM of Oct4A expression standardized to DAPI of three independent experiments. Data is from three independent samples assessed in triplicate. Significance is indicated by **P < 0.01 and ***P < 0.001 as determined by One-Way ANOVA using Dunnett’s Multiple Comparison post-test against the HEY vector control