Fig. 7

Knockdown of KLF6a and KLF7a together but not independently attenuates axon outgrowth from regenerating retinal explants. Morpholino-modified antisense oligonucleotides were delivered to RGCs in vivo via the sectioned optic nerve. Four days later, retinas were isolated, diced, and placed in explant culture for 4 days prior to analysis. Shown are representative images of axon outgrowth. Control morpholino-treated explants exhibit robust axon outgrowth (A) while explants treated with morpholinos targeting either the start codon of both KLF6a and KLF7a (B) or the splice junction of exon 5 of both KLF6a and KLF7a (C) exhibit shorter and fewer axons. (D) Nerve Growth Index for untreated explants (-), explants treated with individual morpholinos (C-MO, control morpholino; 6a, KLF6a-atg; or 7a, KLF7a-atg) or combined treatment to knockdown both KLF6a and KLF7a (6/7atg, KLF6a-atg + KLF7a-atg or 6/7sp, KLF6a-sp + KLF7a-sp). Combined knockdown of KLF6a and KLF7a significantly decreases axon outgrowth compared to control or single morpholino-treated explants. *p < 0.01, ANOVA with Bonferroni post-hoc test. Nerve Growth Index values were normalized to C-MO group that was defined as 100%. Error bars represent SEM.