Fig. 5

Functional endogenous fluorescent tags reveal increased LEA-1 protein expression of multiple isoforms during desiccation. A Endogenous fluorescent tags do not disrupt the function of LEA-1 in forming dauers that are resistant to 1% SDS (dauers formed at 25 °C). B Endogenous fluorescent tags do not disrupt the function of LEA-1 in desiccation survival. lea-1Δ are sensitive to desiccation at 97.5% relative humidity (p < 0.0001, n = 4, unpaired T test) and 60% relative humidity (p = 0.0002, n = 4, unpaired T test). Survival of mNeonGreen tagged LEA-1 worms (mNG::lea-1) and mYPET tagged worms (lea-1::mYPET) was not significantly different from daf-2(e1370) controls after exposure to 97.5% or 60% RH. C Visualization of protein samples extracted from mixed stage cultures, dauer worms, and desiccated worms and blotted with an anti-FLAG antibody indicates multiple isoforms tagged by the two independent tags. The membrane was stripped and blotted for tubulin as a loading control. Total protein in each lane was quantified and normalized to the tubulin loading control. Normalized proteins expression measurements, relative to mixed stage mNG::lea-1 animals, are listed at the bottom of each lane (Norm. Exp.). D A representative dauer worm expressing mNG::lea-1. E A representative desiccated dauer larvae expressing mNG::lea-1. F mNG fluorescence is significantly increased in desiccated worms relative to non-desiccated controls (p = 0.001, n = 3 replicates with 14-15 worms per condition, unpaired T test). G A representative dauer worm expressing lea-1::mYPET. H A desiccated dauer expressing lea-1::mYPET. I Fluorescence intensity is not significantly different between control and desiccated dauer worms expressing lea-1::mYPET (p = 0.39, n = 3 replicates with 15 worms per condition, unpaired T test). Note that all worms were in a daf-2(e1370) background. **p < 0.01, ***p < 0.001