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Fig. 6 | BMC Biology

Fig. 6

From: A tissue-specific protein purification approach in Caenorhabditis elegans identifies novel interaction partners of DLG-1/Discs large

Fig. 6

Confirmation of the DLG-1/ATAD-3 interaction and mapping of the interaction domain. a Western blots showing co-immunoprecipitation (IP) of DGL-1 and ATAD-3. To demonstrate specificity of the detection, samples were treated with control RNAi (GFP RNAi) or RNAi against dlg-1 or atad-3. Left panels: input lysates before IP. Middle panels: DLG-1 IP. Right panels: ATAD-3 IP. b ATAD-3 interacts with DLG-1 in yeast two-hybrid experiments. This interaction depends on the C-terminal ETAV motif of ATAD-3 and the second PDZ domain of DLG-1. Note that ATAD-3 fragments lacking this C-terminal ETAV motif fail to interact with DLG-1 PDZ domains. A fragment containing PDZ 1–3 domains from Drosophila Bazooka was used as a negative control. n.d. not determined. c IP/western blotting experiment shows that, in vivo, the DLG-1/ATAD-3 interaction requires the C-terminal ETAV motif of ATAD-3. Lanes on the left show input signals of DLG-1 and ATAD-3. The lanes on the right show IPs in corresponding lysates. d Progeny produced by N2 animals, or animals expressing full-length ATAD-3 (ATAD-3 FL) and ATAD-3 lacking the C-terminal four amino acids (ATAD ΔETAV) at 15 °C and 25 °C. Total: average total progeny produced. Emb: average number of embryonic lethal progeny produced. Bars represent average values, and error bars the standard deviation. n = 4

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