XB-IMG-170146
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Figure 8. Sustained JAK‐STAT pathway activation reduces neurogenesis. (A) Diagram of the spinal cord graft approach is shown. The caudal portion of the thoracic spinal cord was isolated from transgenic tadpoles (identified by GFP+ eyes and represented as red tadpoles) or their control siblings (black tadpoles), and then it was grafted into WT hosts (black). First heat shock was done 30 min before the spinal cord grafting, and then every 2 days. After 5 days the spinal cord grafts were isolated and analyzed by RT‐qPCR. (B) RT‐qPCR analysis for STAT3 targets (socs3, c‐fosa, and c‐myc), NSPC markers (nestin, vima, and vimb), neurogenic genes (ngn2a and ngn3) and astrocyte marker (aldh1l1). Five biological replicates prepared from different animal pools were analyzed. One sample t‐test with 95% confidence was performed to determine a significant difference (* symbol) from control grafts. (C), (D) Representative animals treated as indicated in (A) were analyzed at 14 days post‐grafting. Longitudinal sections of transgenic (C) and control (D) grafts were analyzed by immunofluorescence. Central canal continuity was analyzed by Sox2/Sox3 (red) and axon continuity by nTubulin (green). In (C) transgenic graft (left side of the figure) is connected (indicated by a dotted line) to the caudal portion of the host spinal cord (right side). The section plane of (C) do not include the connection to the rostral side of the host. In (D) control graft is observed in the middle and connected (dotted lines) to the rostral and caudal portions (left and right sides) of the host spinal cord. White bars indicate 30 μm. The sections depicted are representative of three control and transgenic grafts. Image published in: Tapia VS et al. (2017) © 2017 The Authors. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license
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