Representative images of cerebellar granule neurons transfected with control vector or mycFBXO31 WT plasmid at DIV 0 and analyzed at DIV 3. Arrowheads indicate granule neuron cell bodies. Scale bar equals 50 µm. B. Quantification of percentage of non-polarized granule neurons shown in A. (N = 3, n = 256, mean±SEM, unpaired t-test, **p<0.01). C. Representative images of cultured hippocampal neurons transfected at DIV 1 with control vector, plasmids encoding mycFBXO31 WT or mycFBXO31 ΔF together with the GFP plasmid and immunostained at DIV 7 with α-GFP and α-AnkG antibodies and counterstained with Hoechst. Arrows indicate axon initial segment. Scale bar equals 10 µm. D. Quantification of number of axons in C. A total of 169 cells were analyzed (N = 3, mean±SEM, two-way ANOVA ***p<0.001). E. Representative images of cultured hippocampal neurons from E18 rat embryos transfected with control vector or FBXO31 RNAi#1/CMVGFP plasmid at DIV 1 and immunostained at DIV 6 with α-GFP and α-AnkG antibody and counterstained with Hoechst. Arrows indicate axon initial segment. Scale bar equals 10 µm. F. Quantification of number of axons in E. A total of 121 cells were analyzed (N = 3, mean±SEM, two-way ANOVA ***p<0.001).