Screening for the most effective anti-Xist siRNA. A: Structural illustration of the porcine Xist gene showing the target
site of the 3 designed anti-Xist siRNAs. B: Structural illustration of the plasmid expressing anti-Xist siRNA target
sequences fused with the EGFP gene. The anti-Xist siRNA target site is about 400 bp, composed of partial sequences from exons 1, 5, and 6
of the pig Xist, and covers the target sequences of 3 designed anti-Xist siRNAs. C: Fluorescence microscopy analysis of
EGFP expression, 48 h after co-transfection with anti-Xist siRNA target-EGFP fusion gene expression plasmid and siRNAs into HEK 293 cells.
D: Flow cytometry of relative EGFP+ cell percentage, 48 h after co-transfection with the anti-Xist siRNA target-EGFP fusion gene expression plasmid and
siRNAs into HEK 293 cells. E: Inhibition of Xist expression in female pig fibroblasts by 3 different siRNAs. ** The mean values, calculated
from 3 replicates, are significantly different between the 2 groups (P < 0.01).