Fig. 7

The Cdh23, Harmonin and Myo7aa protein complex is required for proper trafficking of USH2 proteins and can trigger ER stress when defective. (A–O) Immunolabeling of USH2 proteins in (A–C) wild-type (WT) siblings, and (D–F) cdh23, (G–I) ush1c, (J–L) ift88 and (M–O) myo7aa mutants with antibodies against Ush2a (A,D,G,J,M), Gpr98 (B,E,H,K,N) and Whirlinb (C,F,I,L,O). (P) Levels of hspa5 expression based on in situ hybridization technique in whole-mount larvae. For ush1c siblings n=57; ush1c^/ n=51; cdh23 siblings n=35; for cdh23^/ n=33; myo7aa siblings n=24; for myo7aa^/ n=24 analyzed anterior maculae. (Q) Quantification of percentage area per hair cell containing KDEL expression. For ush1c siblings n=85; ush1c^/ n=125; cdh23 siblings n=66; for cdh23^/ n=60; myo7aa siblings n=60; myo7aa^/ n=54 analyzed hair cells for each genotype. (R) Knockdown of cdk5 rescues ER-stress-induced cell death in the ush1c mutant. Bars show average number of TUNEL-positive cells. For ush1c siblings n=52; ush1c siblings + 4 ng cdk5-MO n=18; ush1c^/ n=70; ush1c^/ + 4 ng cdk5-MO n=70 analyzed anterior maculae. Average ± s.d. Statistics were conducted with Student’s t-test. **P<0.01. NS, non significant. Some individual hair cells are outlined by dotted lines. MO, morpholino. Scale bar: 5 µm.