Fig. 5.

Effect of simultaneous silencing of Nor-1, Nurr-1 and Nur-77 on forskolin- or hCG-mediated BeWo cell differentiation. BeWo cells were knocked down for Nor-1, Nurr-1 and Nur-77 simultaneously using siRNA as described in Methods section. To confirm target-specific silencing of all three nuclear orphan receptors, qRT-PCR was performed at 0, 2 and 48 h after forskolin treatment in control siRNA-transfected and silenced cells. The effect of NR4A silencing on BeWo cell fusion and/or hCG secretion was studied after 48 h of forskolin and hCG treatment. a – Comparison of the transcript levels of Nor-1, Nurr-1 and Nur-77 in the control siRNA-transfected and NR4A-silenced cells. Each bar represents relative expression after normalization with 18S rRNA, expressed as means ± s.e.m. of three independent experiments performed in triplicate. b – The fold change in forskolin-mediated BeWo cell fusion in control siRNA-transfected cells and NR4A-silenced cells. Values are shown as means ± s.e.m. of three independent experiments. c – hCG secreted by control and NR4A-silenced cells on treatment with forskolin for 48 h. Data are represented as means ± s.e.m. of three independent experiments performed in duplicate. p ≤ 0.05 is considered statistically significant