Additional file 2.

Confirmation of shRNA-mediated knockdown of endogenous Six3 and Six3OS in developing retina. (A-E) A construct encoding either control hairpin, Six3 targeted hairpin or Six3OS targeted hairpin was electroporated in vivo, into P0.5 retina and harvested at P4.5, and dissociated, and immunocytochemistry against Six3 and GFP, or FISH against Six3OS was performed followed by immunostaining for GFP. GFP-positive cells were counted to analyze the fluorescence intensity for each cell that expressed (A) Six3 or (D) Six3OS. At least 100 GFP-positive cells from three different electroporated retinas were counted for each construct tested. Error bars represent standard error for at least three independent retinas. (A) P < 0.05; (D) P < 0.05. (B-E) Examples of dissociated cells positive for GFP and either Six3 protein or Six3OS RNA.

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Rapicavoli et al. Neural Development 2011 6:32   doi:10.1186/1749-8104-6-32