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. 2013 Aug 12;110(35):14450–14455. doi: 10.1073/pnas.1301588110

Fig. 3.

Fig. 3.

Silencing PKM-ζ leads to the formation of supernumerary axons. (A–F and H–J) E18 rat hippocampal neurons were cotransfected with GFP plasmid and either scrambled (A and B) or PKM-ζ–specific siRNA sequences (PKM-ζ siRNA 1, C and D; PKM-ζ siRNA 2, E and F). Cells were fixed on day 3 in vitro (DIV3) and stained with beta-III-tubulin/Tuj1 (A, C, and E) or Tau and Map2 (B, D, and F). (G–I) Quantitation of the effects of siRNAs on neuronal polarity (G). GFP (H), and pan-aPKC– and aPKC-λ–specific immunofluorescence staining (I) in stage III neurons cotransfected with a GFP plasmid and PKM-ζ siRNA 1. (J) Par3 and Tuj1 immunofluorescence and GFP signal in stage III neurons cotransfected with a GFP plasmid and PKM-ζ siRNA 1. Images in H–J are composites. (Scale bars: 50 μm.) Data are representative images or mean ± SEM of neurons counted (n) from three independent experiments. ***P < 0.001; χ2 test.