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. 2018 Aug 27;115(37):E8746–E8754. doi: 10.1073/pnas.1806933115

Fig. 2.

Fig. 2.

MAP3K LZK promotes axon degeneration by regulating levels of NMNAT2/SCG10. (A) Cas9-expressing DRG sensory neurons were transduced with lentivirus containing sgRNAs to DLK and/or LZK. CRISPR knockout of both DLK and LZK confers additive axon protection in severed distal axons compared with control (Ctrl) sgRNAs or sgRNAs targeting DLK (sgDKL) or LZK (sgLZK). A repeated-measures ANOVA was performed with post hoc Bonferroni tests for multiple comparisons, where ****P < 0.001 (n = 3). (B) Bright-field images of distal axons at the indicated time points after axotomy. (Scale bar, 5 μm.) (C) Western blots of phosphorylated MKK4 and JNK from axon-only extracts in Cas9-expressing DRGs transduced with Ctrl sgRNAs or sgDLK and sgLZK. Western blots are representative of at least three independent experiments. (D) Western blots of NMNAT2 and SCG10 from axon-only extracts derived from DRG sensory neurons transduced with the indicated sgRNAs. (E) Quantification of NMNAT2 and SCG10 protein levels in the presence of the indicated sgRNAs [***P < 0.001, single-factor ANOVA with post hoc t tests (n = 3)]. Error bars represent SEM.