DAF-15:mCherry expression during C. elegans development
(A) A cartoon diagram of the DAF-15::mCherry CRISPR/Cas9 construct used in this study.
(B and C) Western analysis and representative microscopy images of mixed population samples of control (N2 WT) and cas9 knock-in (DAF-15::mCherry) worms. Probing with anti-mCherry revealed a single band (B) corresponding to DAF-15::mCherry at ∼250kD, and a visible red signal in the head region (C). Scale bar is 100um.
(D and E) Western analysis and representative microscopy images of DAF-15::mCherry worms fed either HT115 or daf-15(RNAi) (arrested at L3 stage). The single band (D) detected in the control sample was strongly reduced by daf-15(RNAi); the visible red signal detected in the head region of the control sample was strongly reduced by daf-15(RNAi). Scale bar is 100um.
(F) Microscopy images and western analysis of synchronized, staged populations of DAF-15::mCherry worms revealed the DAF-15:mCherry level decreased with age.