FIGURE

Fig. 3

ID

ZDB-FIG-220715-51

Publication

Berger et al., 2022 - Mob4-dependent STRIPAK involves the chaperonin TRiC to coordinate myofibril and microtubule network growth

Other Figures

All Figure Page

Back to All Figure Page

Fig. 3

Mob4 is located at the Z-disk, where it might be involved in the regulation of myofibril assembly.

(A) At 3dpf, antibodies against human MOB4 colocalised with antibodies against the Z-disc protein α-Actinin (n = 6 per genotype). (B) At 3 dpf, the birefringence of mob4^geh homozygotes was significantly higher in the transgenic background of Tg(cry:GFP;-503unc:mob4). Also compared to non-transgenic siblings (100 ± 2%), the birefringence of Tg(cry:GFP;-503unc:mob4) transgenic mob4^geh homozygotes (108 ± 1%) and siblings (106 ± 1%) was significantly higher. Crosses represent averaged birefringence of clutches with a minimum of 4 larvae per genotype (n = 5 clutches). Data are presented as mean ± SEM; *** P < 0.001 and ** P < 0.01 by one-way ANOVA with post hoc Tukey’s test. (C) In 3-dpf-old siblings and mob4^geh homozygotes, Mob4-GFP fusion protein (green) expressed via Tg(cry:GFP;-503unc:mob4-GFP) colocalised to t-tubules (red, arrowhead) marked in the Tg(acta1:mCherryCAAX) transgenic background (n = 3 per genotype). (D) Highlighting F-actin with transgenic Tg(acta1:lifeact-GFP) in green confirmed residual myofibril striation and revealed disorganised thin filaments within mob4^geh homozygotes at 3 dpf. Sarcolemma and t- tubules were labelled by mCherry fluorescence (red) in the Tg(acta1:mCherryCAAX) transgenic background (n = 6 per genotype). Boxed areas are magnified. (E) Labelling of F-actin with phalloidin revealed that the robust myofibril striation of siblings was reduced in mob4^geh homozygotes at 3 dpf (n = 6 per genotype). (F) At 3 dpf, antibodies against α-Actinin that mark sarcomere’s Z-disks showed the typical striation of the myofibril in siblings and mob4^geh homozygotes (n = 4 per genotype). (G) Visualisation of the cephalic muscles in the transgenic Tg(−503unc:GFP) background revealed that, in contrast to siblings, a gap was formed between the two hyohyoideus (hh) muscles in mob4^geh homozygotes at 3 dpf (representative Z-stacks) (n = 3 per genotype). (H) At 6 dpf, representative Z-stack projections of Alcian blue stained larvae depicted cartilage malformations in mob4^geh homozygotes and a widened angle formed by the two ceratohyal cartilage structures (dotted lines) (n = 4 per genotype). Designations: ceratohyal (ch); Meckel’s cartilage (m); palatoquadrate (pq). Scale bar sizes are indicated.

Expression Data

Genes:	EGFP GFP mCherry mob4
Antibodies:	Ab1-actn Ab1-mob4
Fish:	TU mob4^pc43/pc43 (TU) pc21Tg; pc22Tg mob4^pc43/pc43; pc21Tg; pc22Tg pc22Tg; pc47Tg mob4^pc43/pc43; pc22Tg; pc47Tg
Anatomical Terms:	skeletal muscle cell filamentous actin skeletal muscle cell sarcolemma skeletal muscle cell striated muscle thin filament skeletal muscle cell T-tubule skeletal muscle cell Z disc
Stage:	Protruding-mouth

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	mob4^pc43/pc43 mob4^pc43/pc43 (TU) mob4^pc43/pc43; pc21Tg; pc22Tg mob4^pc43/pc43; pc46Tg
Observed In:	ceratohyal cartilage left side ceratohyal cartilage right side hyohyoideus left side hyohyoideus right side myotome skeletal muscle myofibril skeletal muscle cell striated muscle thin filament
Stage Range:	Protruding-mouth to Day 6

Phenotype Detail

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 @ PLoS Genet.