Fig. 7. CeAJ proteins are mislocalised differently in epidermal cells of let-413(RNAi) and dlg-1(RNAi) embryos. Embryos were stained with anti-HMP-1 or anti-JAM-1 along with anti-UNC-70 antibodies and DAPI. Confocal sections were captured and analysed as described in Materials and Methods. (A) Bar graph showing the spread of CeAJ proteins (JAM-1, pink bars, and HMP-1, orange bars) along the lateral membrane of epidermal cells in mature wild-type (mWT), dlg-1(RNAi), let-413(RNAi) and early wild-type (eWT) embryos. Values are means ± s.e.m. (B) Bar graphs showing the position of the same CeAJ proteins along the lateral membranes of epidermal cells. The graphs show the percentage of membranes where JAM-1 or HMP-1 were detected at the top (T), middle (M) or bottom (B) of the lateral membrane (see C). Proteins were often distributed over more than one membrane section and so the total percentage for each category is greater than 100%. Note that the spread and position of HMP-1 was not determined in early wild-type embryos (ND in graphs). The numbers of lateral membranes scored for each set of embryos were as follows. Wild-type: 71 (JAM-1), 88 (HMP-1); dlg-1(RNAi): 57 (JAM-1), 80 (HMP-1); let-413(RNAi): 78 (JAM-1), 85 (HMP-1); early wild-type: 85 (JAM-1). (C) Schematic diagram of a transverse section through the mature epidermal layer of a wild-type embryo (see tan bracket, Fig. 6A), indicating how a lateral membrane was sectioned to score protein position. Nuclei are shown in blue, CeAJs in pink and lateral membranes in green. (D-I) Z-axis rotations showing transverse sections through the epidermal layer of mature wild-type (D,G), dlg-1(RNAi) (E, H), and let-413(RNAi) (F,I) embryos after immunostaining with MH27 (red) (D-F) or anti-HMP-1 (red) (G-I), along with anti-UNC-70 (green) antibodies and DAPI (blue). The basal extent of UNC-70 denotes the position of the epidermal basal membrane and the nucleus of the cell under investigation is indicated with an asterisk, as in C. (J,K) Internal view of a let-413(RNAi) dlg-1::gfp transgenic embryo after immunostaining with anti-GFP antibody alone (J), and both anti-GFP and MH27 antibodies (K), showing that DLG-1 is mislocalised in the absence of LET-413 but remains colocalised with JAM-1. (L,M) Internal view of a wild-type (L) and a dlg-1(RNAi) (M) let-413::gfp transgenic embryo after immunostaining with an anti-GFP antibody, illustrating that LET-413 is not mislocalised in the absence of DLG-1. Scale bars, 5 µm (D-I); 10 µm (J-M).