IMAGE

Fig. 2

ID
ZDB-IMAGE-111215-34
Source
Figures for Phillips et al., 2011
Image
Figure Caption

Fig. 2 Mutation and morpholino knockdown of ush1c produce behavioral defects. (A) RT-PCR of control and splice-blocking morpholino-injected embryos from 1 to 5 dpf reveals a smaller splice form lacking exon 2 that predominates for the first 4 days of development and persists through the fifth day. (B) Western blots from extracts of control + ush1c MO-injected embryos from 3 to 5 dpf (left) and ush1cfh293 homozygous mutant embryos alongside wild-type siblings (right) confirm morpholino efficacy and antibody specificity. Samples from 3 dpf to 5 dpf control larvae probed with the zebrafish harmonin antibody show a 60 kDa band consistent with the predicted size of isoform A. Additional smaller bands are visible faintly in the 4 dpf control and are more pronounced and abundant in the 5 dpf control. The band corresponding to the isoform A variant is undetectable in 3 dpf and 4 dpf morphant samples. The major band reappears at 5 dpf at a slightly smaller weight, and there is a reduced number of smaller bands in this sample. In extracts from 5 dpf ush1cfh293 homozygous mutants, no bands are observed. Extracts from wild-type siblings from the same clutch show the characteristic banding pattern of the 5 dpf uninjected controls, allowing for the slightly longer running time for this gel. A pan-actin antibody was used as a loading control. (C) Behavioral assays of control, mutant and morpholino-injected larvae show (top) impaired startle response and balance, and (bottom) reduced optokinetic response, tracking an average of 50% or fewer of the dark stripes (average targets tracked for wild type: 11.5±1.27; ush1c mutants: 6.13±2.78, P<0.0001; ush1c MO: 5.13±2.85, P<0.0001; n=30 for each group).

Figure Data
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Dis. Model. Mech.