ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Casq mRNA in selected tissues from adult zebrafish (A). The levels of RNA expression are relative to two different housekeeping genes, beta-2-microglobulin (B2M) and elongation factor1-α (EF1α). Mean values and standard error were determined from triplicate runs of the qPCR assay in three different tissue preparations. (B). Anti-Casq antibodies reactivity on total homogenates (40 μg per lane) obtained from pooled (N = 3) skeletal muscle, heart, and brain of adult zebrafish. In the control lanes, (anti-rabbit) blot incubation was performed in the presence of anti-rabbit conjugate with alkaline phosphatase only. Molecular mass markers (in kDa) are indicated. (C)In vitro deglycosylation of Casqs. Immunodetection of Casq by CC antibody in membrane-enriched fraction P4 (see “Materials and Methods” section) after endoglycosidase F digestion, with 10 μg loaded per lane.

Adult brain subcellular fractionation: immunochemical profile. Immunoblot analysis of brain subcellular fractions. Equal protein amounts (40 μg) from each fraction were analyzed with CC antibodies to recognize Casqs and with antibodies specific for synaptotagmin1 (SYT1), calreticulin, NDUFS3 (as described in “Materials and Methods” section). Raw data derived from densitometric analysis are presented in Supplementary Table S1.

Identification of Casqs in adult brain fractions. (A) Relative abundance of the two detected isoforms. (B) Mapping of peptides identified by mass spectrometry on Casq sequences. In red is the consensus site for N-glycosylation.

In situ analysis of casq1a [(A–D) and casq2 panels (E–H)] transcripts in sagittal brain sections (A,E, rostral to the right). Whole-brain sections and control sections are presented in Supplementary Figure S1. Anatomic terminology was inferred from Ullmann et al. (2010). CCe, corpus cerebelli; GCL, granule cell layer; Lca, caudal lobe of cerebellum; ML, molecular layer; PCL, Purkinje cell layer; PVZ, periventricular zone of optic tectum; TeO, optic tectum. (A,E) Bar: 200 μm. (B–D,F–H) Bar: 20 μm.

Immunofluorescence staining of sagittal brain sections area restricted to the cerebellum and the optic tectum (rostral to the right) decorated with CC (A,C–E) and MC (F,H–K) antibodies. (B,G) Images collected from control sections (see “Results” section) and acquired with the same conditions as in (A) and (F), respectively. (A,B,F,G) Epi-fluorescence signals. (C–E,H–K) Images obtained by single optic section of confocal analysis. The box in (C) indicates the area shown at higher magnification in (E). (D) PCL in a different region and focal plane. Arrows: axons of Purkinje cells. The abbreviations are the same as in Figure 4. (A,C,F,H,I) Bar: 100 μm. (E) Bar: 25 μm. (D,J,K) Bar: 7.5 μm.

Co-staining of cerebellum with Casq and neuron-specific markers. (A–C) Epi-fluoresence signal obtained by double immunofluorescence with CC antibody (green) and anti-parvalbunin antibodies (red) in the cerebellum area. (C) A merge of the two images. Arrowheads indicate the axons of Purkinje cells. (D–F) Epi-fluoresence images obtained by double-staining with MC antibody (green) and anti-parvalbunin antibodies (red). (D) A merge of the two images. (G–I) Confocal images obtained by double-staining with MC antibody (green) and anti-NeuroD1 antibody (red). (J–L) Higher magnification of granule cells in the central area of the cerebellum double-labeled with MC (green) and anti-NeuroD1 antibody (red). Bar: 25 μm.