Skip to main content
. Author manuscript; available in PMC: 2020 Apr 13.
Published in final edited form as: Nat Neurosci. 2019 Jan 14;22(2):167–179. doi: 10.1038/s41593-018-0300-4

Figure 6. STMN2 mutant neurons have neurite outgrowth and regrowth deficits similar to neurons treated with siTDP43.

Figure 6

(a) Knockout strategy targeting two constitutive exons of the human STMN2 gene. (b-d) STMN2 elimination was confirmed in the HUES3 Hb9::GFP line by RT-PCR analysis of genomic DNA (b), by immunoblot analysis (c), and by immunofluorescence (d). Similar results were obtained from n=2 biologically independent experiments. (e) Experimental strategy used to assess the cellular effect of STMN2 elimination in hMNs. (f-h) Sholl analysis of hMNs with and without STMN2 and in the absence (g) or presence (h) of an ROCK inhibitor (Y-27632, 10 μM). Lines represent sample means and shading represent the SEM with unpaired t-test between siTDP43 and siSCR, two-sided, P value < 0.05 with all values in Supplementary Table 1. Similar results were obtained in n=2 biologically independent experiments. (i) Experimental strategy used to assess the cellular effect of STMN2 elimination in hMNs after axonal injury. (j-k) Axonal regrowth after injury. Representative micrographs of hMNs in the microfluidics device prior to and after axotomy (j). Measurements of axonal regeneration after axotomy. Individual neurites are displayed as dots along with the mean and SD (Unpaired t-test, two-sided, P value < 0.05 18hrs=0.0005, 24hrs=0.0001, 48hrs=<0.0001) (k). Similar results were obtained in n=4 devices from 2 independent experiments. Similar results were obtained in n=2 biologically independent experiments.