Fig 1. Expression of ALG-1 and ALG-2 during adulthood. (A) Western blot of FLAG::GFP-tagged ALG-1 in protein samples from L4 and adultstage transgenic animals. Tubulin levels serve as a loading control. (B) Western blot of FLAG::RFP-tagged ALG-2 in protein samples from L4 andadult stage transgenic animals. A sample from L4 stage non-transgenic (nt) animals demonstrates the specificity of the anti-FLAG antibody.Tubulin levels serve as a loading control. (C) Expression of endogenous ALG-1 tagged with GFP in L4, 2d and 5d adult animals visualized byfluorescence microscopy. The panels of non-transgenic animals show that auto-fluorescence contributes to the signal detected in the intestine ofadults. Micrographs were captured at 40x magnification with 26 ms exposure. (D) Expression of endogenous ALG-2 tagged with RFP in L4, 2d
and5d adult animals visualized by fluorescence microscopy. The panels of non-transgenic animals show that auto-fluorescence contributes to thesignal detected in the intestine of adults. Micrographs were captured at 40x magnification with 450 ms exposure. (E) Simultaneous expression ofGFP::ALG-1 and RFP::ALG-2 in L4 and 5d adult animals visualized by fluorescence microscopy. Micrographs were captured at 40x magnificationwith equivalent exposures at L4 and 5d adult stages. (F) qRT-PCR analyses of
alg-1 and
alg-2 mRNA levels in adults relative to L4 stage in the
spe-9(
hc88)strain, averaged from three independent experiments. The sterile
spe-9 background was used to avoid potential signal from progeny animals.The error bars represent SEMs. P<0.01, P<0.001 (t-test).