Anti-REST antibody injection and efficient knockdown of REST by REST-LNA.
A and A′, immunostaining of anti-REST antibody injected oocytes only by secondary antibody; B and B′, immunostaining of non-injected oocytes only by secondary antibody; C and C′, immunostaining of oocytes post-2 h of anti-REST antibody injection only by secondary antibody. FITC-labeled donkey anti-rabbit IgG was used as secondary antibody, which was used to immunostain anti-REST antibody. Green, anti-REST antibody; blue, DNA. Scale bar, 50 μm. D, effectively match of anti-REST antibody to the porcine REST among whole oocyte proteins was verified by Western blotting. E, efficient knockdown of REST mRNA by REST-LNA in porcine oocytes checked by Q-PCR. Significant difference was found between the two groups by Student's t test (p < 0.001). F, efficient knockdown of REST protein in porcine oocytes checked by Western blotting analysis. CON, MII oocytes; REST-LNA, REST-LNA injected MII oocytes.