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. Author manuscript; available in PMC: 2021 Feb 1.
Published in final edited form as: Mol Neurobiol. 2019 Nov 7;57(2):1146–1158. doi: 10.1007/s12035-019-01796-2

Figure 5. Oligomycin and forskolin promote loss of NMNAT2 from axons.

Figure 5.

(A) Neurons were treated with forskolin, oligomycin, or a combination of forksolin and oligomycin for 2 hr and axonal NMNAT2 levels were assessed. Shown below is quantification of NMNAT2 levels as a ratio of vehicle control (N=4, ****p<0.0001, *p=.0147, 0.103 respectively with single factor ANOVA with Bonferroni post hoc T-test correction). (B) Neurons treated as in (A) were also assessed for axonal SCG10 levels with quantification below (N=3, ****p<0.0001, **p= 0.0076, and *p=0.0158 from single factor ANOVA with Bonferroni post hoc T-test correction). (C) Oligomycin reduces axonal NMNAT2-myc protein expressed via lentivirus from ubiquitin promoter. Neurons were treated with vehicle or oligomycin (500nM) for two hours. Quantification of NMNAT2-myc levels is shown to the right (N=4 *p=0.0144 unpaired t-test). (D, E) Axonal NMNAT2 protein is reduced by oligomycin in the presence of Cas9 and sgRNAs to DLK/ LZK (N=5, ***p=0.0011 and 0.0064 for unpaired T-tests within sgCtrl and sgDLK/LZK respectively). Error bars represent +/-1 SEM.