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. 2019 Aug 22;8(9):956. doi: 10.3390/cells8090956

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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The effect of combined siGSK3β + siRTP801 on GSK3β and RTP801 immunoreactivity in RGC in vitro. (A) GSK3β and RTP801 mRNA levels in cultured adult rat retinal cells after transfection with the optimal concentrations of siGSK3β (25 nM each) to confirm significant GSK3β and RTP801 knockdown. Lipo = Lipofectamine 2000 (*** = p < 0.001). Both (B) RTP801 and (C) GSK3β⁺ immunoreactivity was present in RGC from sNBA and siEGFP control cultures, whilst treatment with combined siGSK3β + siRTP801 abolished GSK3β and RTP801 immunoreactivity in RGC. (Scale bar in B and C = 20 μm). Combined siGSK3β + siRTP801 significantly increased (D) the number of surviving βIII-tubulin⁺ RGC, (E) % of RGC with neurites and (F) the mean longest neurite length. (G) GSK3β⁺ and RTP801⁺ immunoreactivity was present in sNBA and siEGFP control cultures, whilst treatment with combined siGSK3β + siRTP801 significantly reduced RTP801 and GSK3β immunoreactivity in RGC. * = p < 0.05; **p < 0.01; *** = p < 0.001). n = 2 wells/treatment, 3 independent repeats (total n = 6 wells/treatment).