36K is mainly expressed in zebrafish CNS. (a) Exon‐exon junction spanning rt‐PCR showing expression of 36K in different organs of adult zebrafish, with ef1a as positive control. (b) rt‐PCR showing 36K expression in zfl during development. Whole larvae were used to prepare RNA at different dpf. At 6 dpf, additionally, the complete head region was separately taken for RT PCR depicted as 6 dpf head (Lane 7). (c) Quantitative rt‐PCR analysis of 36K expression during zfl development normalized to β‐actin and ef1a depicting calibrated normalized relative quantities (CNRQ) indicated an increase in 36K transcript expression from 3 dpf. (d) Western blot against 36K with whole larvae at different developmental stages, adult brain and spinal cord with β‐actin as loading control accordingly. (e) Anti‐36K staining in sagittal paraffin section from adult zebrafish showing specific expression in brain and spinal cord. Sections were counter stained with hematoxylin. Scale bar: 2 mm. The red inlay zooms in to show specific expression in brain. Scale bar: 200 μm. (f) Anti‐36K fluorescent staining in transverse paraffin section from adult zebrafish showing specific expression in spinal cord. Dashed lines depict fish skin. Scale bar: 200 μm. The green inlay zooms onto anti‐36K fluorescent staining in the spinal cord. Scale bar: 40 μm. (g) Anti‐36K (red) and anti‐EGFP (green) fluorescent staining in sagittal paraffin section from 3 dpf Tg(olig2:GFP) zfl hindbrain and spinal cord. A merge of the two labels is shown on the right, where the asterisk marks co‐localization (in yellow) of the olig2‐GFP and 36K protein expression in the myelin fibers of the spinal cord. Scale bar: 200 μm
|
