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. 2012 Mar 1;26(5):461–473. doi: 10.1101/gad.182998.111

Figure 6.

Figure 6.

Amd1 is required for ESC maintenance. (A) ESCs transfected with negative control mimics or with miR-762 mimics with or without the pTracer-Amd1 vector. Cells were stained with AP 4 d after transfection. (B) qRT–PCR analysis of mRNAs 4 d after transfection as in A. (C) Western blot analysis showing Amd1 and ESC markers 4 d after transfection with miR-762 mimics. GAPDH was used as loading control. (D) qRT–PCR showing Amd1 knockdown with siRNAs. Scrambled siRNAs are shown as a control. (E) AP staining of ESCs transfected with siRNAs to Amd1. (F) qRT–PCR showing mRNA expression levels following transfection of ESCs with siRNAs targeting Amd1 with and without the addition of spermine (SPM). P-values show significant differences between ESCs and siAmd1 samples or between siAmd1 plus and minus spermine. (G) Western blot showing Amd1, Oct4, and Sox2 levels following transfection of ESCs with siRNAs targeting Amd1. GAPDH is shown as a loading control. (H) Immunofluorescence staining for ESC marker Oct4 in ESCs transfected with pools of siRNA targeting Amd1 or scrambled siRNA. Bars, 20 μm. Values are means ± SD. (*) P < 0.05.