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Figure 4


Fig. 4. Cellular localization and functional rescue of calsequestrin. (A,B) Wild-type worms stained with (A) anti-CSQ-1 antibody and (B) anti-UNC-68 (RyR) antibody show punctate and mesh-like staining in body-wall muscles. (C) The csq-1(jh109) mutant stained with anti-UNC-68 antibody shows similar punctuate and mesh-like staining pattern. (D) The unc-68(e540) mutant stained with anti-CSQ-1 antibody show a dispersed punctate pattern; no mesh-like pattern was observed. (E) csq-1-null mutant csq-1(jh109) transformed by constructs encoding WT, {Delta}N23-31 and Bedouin CSQ-1 show mesh-like pattern. (F) {Delta}N32-43 shows aggregated pattern. (G) K111A shows diffused pattern. (H) {Delta}C17 and {Delta}C26 show dispersed pattern similar to that shown in D. (I) Survival rates of wild-type and transgenic animals expressing mutated CSQ-1 under high-[Ca2+] and low-[Ca2+] conditions. Young F1 transgenic worms were grown on NGM plates with 20 mM EGTA and 20 mM CaAc. Survival rates of worms with rescued CSQ-1 function was compared with that of wild-type animals at 20°C. Over 50 animals were tested for each data point of a single set of experiments, and each experiment was repeated three times.





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