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Fig. 4. NRX IV, NRG and CONT are interdependent for their localization at the SJs. (A-D) This region of the wild-type epidermis at stage 16 shows the SJ localization of NRX IV (A, blue), CONT (B, red) and NRG (C, green), and the merged image (D). (E-H) Epidermis from a nrx IV⁴³⁰⁴/nrx IV⁴³⁰⁴ embryo at stage 16 with a loss of NRX IV protein (E, blue), a diffuse localization of CONT around the cell cortex (F, red) and a uniform basolateral localization of NRG (G, green). White arrowheads indicate the basolateral distribution of NRG (compare G with C). CONT is also seen in intracellular vesicles, as indicated by red arrowheads in F and H. (I-L) This part of the epidermis from a nrg¹/Y embryo at stage 16 shows the loss of NRG staining (I, blue). White arrowheads indicate the basolateral distribution of both NRX IV (J, red) and CONT (K, green). Compare the merged image L with D. (M-P) Part of the epidermal epithelial layer from homozygous cont^ex956 mutant embryo at stage 16, showing total loss of CONT (M, blue) and mislocalization of NRX IV (N, red) and NRG (O, green) along the basolateral area, as indicated with arrowheads (N-P). Merged image in P. (Q-V) Epidermis from hs-cont/hs-cont;cont^ex956/cont^ex956 homozygous embryos at stage 16 heat-shocked to induce the expression of CONT (Q and T, blue), show that NRX IV (R, green) or NRG (U, green) localization at SJs has been restored (compare arrowheads in R with N or in U with O). Merged images are shown in S and V. (W-Z) Epidermis from cont^ex956/cont^ex956 embryo rescued by SpeI genomic fragment at stage 16 shows normal expression and localization of CONT (W), and restoration of NRX IV (X) and NRG (Y) to SJs. A merged image of these proteins is shown in Z. Arrowheads indicate localization of these proteins at SJs. Scale bar: 16 µm.

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