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. 2020 Jun 25;9:e56935. doi: 10.7554/eLife.56935

Figure 2. The NG2-DsRed/S100β-GFP mouse line can be used to reliably identify PSCs in healthy and stressed NMJs.

Figure 2.

(A–B) Representative images of NMJs identified by fBTX labeled nAChRs from S100β-GFP (A) and S100β-GFP;NG2-dsRed (B) EDL. (C–D) The co-expression of GFP and dsRed has no discernible negative effects on NMJ fragmentation or PSC number in the EDL muscle of young adult mice. (C) The average number of PSCs per NMJ is unchanged between S100B-GFP mice and S100β-GFP;NG2-dsRed mice. (D) The average number of nAChR fragments per NMJ, as determined by analysis of continuity of fBTX labeled nAChRs, is unchanged between wild-type, S100B-GFP, and S100β-GFP;NG2-dsRed animals. (E–H) PSCs in stressed muscle co-express S100β-GFP and NG2-dsRed. Representative images of NMJs identified by fBTX labeled nAChRs in S100β-GFP;NG2-dsRed mice shows co-expression of S100β-GFP and NG2-dsRed by PSCs in healthy uninjured (E), at 4d (F), and 7d (G) post-fibular nerve crush, and in P120 SOD1G93A (H) EDL. At non-synaptic sites, GFP-positive cells do not express dsRed (hollow arrow; E, F, G, H) and dsRed-positive cells do not express GFP (filled arrow; E, F, G, H). Error bar = standard error of the mean. Scale bar = 10 μm (A–B), 50 μm (E–H), and 12.5 μm (E’-H’ and E’=H’).