rad1 a1047 antibody  (ABclonal Biotechnology)

Journal: Cell Research

Article Title: MIWI and piRNA-mediated cleavage of messenger RNAs in mouse testes

doi: 10.1038/cr.2015.4

Figure Lengend Snippet: Temporal regulation of the piRNA target genes Xrcc2 and Rad1 is essential for sperm formation. (A) Dual-luciferase reporter assays to determine the effects of piR-mmu-170840 on the Xrcc2 reporter (left), and piR-mmu-604682 on the Rad1 reporter (right) in transfected GC-2spd(ts) cells. A mutant piRNA (Mut) served as negative control in each case. (B) Western blot analyses of Xrcc2 and Rad1 expression in enriched SC and RS, with β-actin serving as the loading control. (C , D) qRT-PCR (C) or northern blot (D) analyses of the mRNA levels of Xrcc2 and Rad1 in Miwi +/− and Miwi −/− mouse spermatogenic cells, with GAPDH serving as an internal control. Mean ± SEM of three separate experiments were plotted. Student's t -test significance: * P < 0.05, ** P < 0.01, *** P < 0.001. (E) PCR analyses of GFP DNA in extracted genomic DNA from late spermatids (top) and epididymal sperms (bottom) from mouse testes transduced with IRES-EGFP (lanes 4-8), Xrcc2-IRES-EGFP (lanes 10-14), or Rad1-IRES-EGFP (lanes 16-20), respectively. Genomic DNA samples from Oct4-GFP transgenic mice (lane 1) and wild-type background mice (lane 2) served as positive and negative controls, respectively. Results shown are representative of three independent experiments.

Article Snippet: Rabbit anti-Rad1 polyclonal antibodies (A1047) was from ABclonal Technology and anti-GFP (MBL-598) was from MBL.

Techniques: Luciferase, Transfection, Mutagenesis, Negative Control, Western Blot, Expressing, Control, Quantitative RT-PCR, Northern Blot, Transduction, Transgenic Assay