FIGURE 4: Expression and subcellular localization of CPL-1::mChOint and its rescuing activity. (A) Schematic representation of the fosmid reporter of the
cpl-1 gene fused in frame with mChOint. Solid boxes indicate exons, and wavy lines indicate introns. The red box indicates the mChOint ORF. (B) Representative images showing the rescue of embryonic lethality of
cpl-1(
ok360) mutants by the transgenic array yqEx620 expressing the
cpl-1 fosmid reporter (a-c) DIC images showing
cpl-1(
ok360) embryos that arrested at early embryonic stages. (d-f) DIC and fluorescence images of
cpl-1(
ok360) mutants carrying yqEx620, which developed normally. Bars, 5 1/4 m. (C) Quantification of the rescuing effect of yqEx620 on the embryonic lethality of
cpl-1(
ok360) mutants grown at 20 degrees C. We analyzed 100 embryos for each genotype, and data were derived for three repeats. ***p < 0.001, unpaired t test. (D) Expression and subcellular localization of CPL-1::mChOint driven by the
cpl-1 promoter. Images of DIC, CPL-1::mChOint, and LAAT-1::GFP (driven by the
ced-1 promoter) and merged images of CPL-1::mChOint with LAAT-1::GFP are shown for a 1.5-fold embryo, an L1 larvae, the germ line, and the hypodermis. Bars, 5 1/4 m. Arrowheads indicate some of the foci with colocalization of CPL-1::mChOint and LAAT-1::GFP. (E) Quantification of the colocalization of LAAT-1::GFP driven by the
ced-1 promoter and CPL-1::mChOint driven by the
cpl-1 promoter. (F) Rescuing effect of CPL-1::mChOint transgenic arrays yqEx620 (
cpl-1 fosmid reporter), yqEx688 (Pcpl-1cpl-1::mChOint), yqEx690 (Pcpl-1cpl-1
(del51)::mChOint), yqEx743 (Pcpl-1cpl-1(C144A)::mChOint), and yqEx746 (Pcpl-1cpl-1(G230R)::mChOint) on cell corpse phenotype in
cpl-1(
yq89)mutants.
cpl-1 mutants carrying transgenic arrays were grown at 25 degrees C, and cell corpses were scored in animals at 24 h post L4 (n=15 for each genotype). Comparisons were performed between N2 and
cpl-1(
yq89) mutants with or without transgenic arrays as indicated using an unpaired t test. ***p < 0.001.