Figure 1

Effects of overexpressing SLIT2 on the expression of the ROBO1, ROBO2, ROBO3 and ROBO4 genes. The granulosa cells were transfected with a reconstructed pYr-adshuttle-4-SLIT2 vector, a pYr-adshuttle-4 empty vector (negative control) or no plasmid (blank control). (A) The expression of the SLIT2 gene before and after the GCs were transfected with the pYr-adshuttle-4-SLIT2 expression vector for 24 h was examined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values on the bar graphs represent the mean ± SEM of 10 hens (n = 10) from a representative experiment. (B) The expression levels of the SLIT2 protein in the GCs before and after the transfection with the pYr-adshuttle-4-SLIT2 vector were detected by western blotting. β-actin was used as a loading control. The blots were cropped, and the gels were run under the same experimental conditions. (C) The influence of the SLIT2 overexpression on the ROBO1, ROBO2, ROBO3 and ROBO4 mRNA abundance in the granulosa cells from the prehierarchical follicles (6 to 8 mm in diameter) was examined. (D) The effects of the SLIT2 overexpression on the protein levels of ROBO1 and ROBO2. For each group, the superscript symbol above the bar indicates that the difference was significant compared to the control group **P < 0.01, *P < 0.05.