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. 2018 Oct 12;7:e38309. doi: 10.7554/eLife.38309

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© 2018, Medina-Cano et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Figure 7. — (A) Neurite number per GC body cluster across 21 hr in laminin and poly-D-lysine-coated wells as measured with Incucyte live cell imaging system (Control, n = 3; En1-Cre; Srd5a3^fl/-, n = 3). (B) Representative image of control and En1-Cre; Srd5a3^fl/- GCs after 21 hr in laminin and poly-D-lysine-coated wells. Scale bar 100 μm. (C) Neurite number normalized to GC body clusters after 21 hr with laminin/poly-D-lysine coatings, fibronectin or IgSF-CAMs-coated wells (NrCAM and L1CAM). Each dot per coating represents results from a single mutant mouse. All GC cultures were performed in technical triplicates (Control, n = 3; En1-Cre; Srd5a3^fl/-, n = 3). (D) Neurite number and number of migrating neurons in GC re-aggregates plated for 24 hr on laminin and poly-D-lysine and (E) L1CAM coated surfaces. Five aggregates were quantified per mouse and coating condition (Control n = 5, En1Cre; Srd5a3^fl/-, n = 7). (F) Representative electron microscopy images of cerebellar ML sagittal view at P21 taken from control (n = 3) and Srd5a3 mutant (En1-Cre; Srd5a3^fl/- n = 1; Atoh1-Cre; Srd5a3^fl/- n = 2) mice. Scale bar 2 μm. Parallel fibers show a single orientation in control ML, whereas some exhibit an abnormal perpendicular orientation in the most outer ML in the mutant mice (G, red arrowhead). Two-tailed Student t-test was used for statistics. *p<0.05; **p<0.01. Results are presented as mean ± s.d.

Figure 7—figure supplement 1. — (A) Neurite number per GC body cluster across 21 hr in laminin and poly-D-lysine-coated wells as measured with Incucyte live cell imaging system (Control, n = 3; En1-Cre; Srd5a3^fl/-, n = 3). (B) Representative image of control and En1-Cre; Srd5a3^fl/- GCs after 21 hr in laminin and poly-D-lysine-coated wells. Scale bar 100 μm. (C) Neurite number normalized to GC body clusters after 21 hr with laminin/poly-D-lysine coatings, fibronectin or IgSF-CAMs-coated wells (NrCAM and L1CAM). Each dot per coating represents results from a single mutant mouse. All GC cultures were performed in technical triplicates (Control, n = 3; En1-Cre; Srd5a3^fl/-, n = 3). (D) Neurite number and number of migrating neurons in GC re-aggregates plated for 24 hr on laminin and poly-D-lysine and (E) L1CAM coated surfaces. Five aggregates were quantified per mouse and coating condition (Control n = 5, En1Cre; Srd5a3^fl/-, n = 7). (F) Representative electron microscopy images of cerebellar ML sagittal view at P21 taken from control (n = 3) and Srd5a3 mutant (En1-Cre; Srd5a3^fl/- n = 1; Atoh1-Cre; Srd5a3^fl/- n = 2) mice. Scale bar 2 μm. Parallel fibers show a single orientation in control ML, whereas some exhibit an abnormal perpendicular orientation in the most outer ML in the mutant mice (G, red arrowhead). Two-tailed Student t-test was used for statistics. *p<0.05; **p<0.01. Results are presented as mean ± s.d.