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. 2022 Nov 20;22(1):e13742. doi: 10.1111/acel.13742

FIGURE 1.

FIGURE 1

Multiplex screening reveals bet‐1 is involved in regulation of actin. (a) Schematic of cross‐species screening. CRISPR‐Cas9 screening was performed in BJ fibroblasts with 0.1 μM cytochalasin D and top hits were screened in Caenorhabditis elegans with 90% candidate RNAi and 10% act‐1 RNAi. (b) Images of N2 animals grown on 90% candidate RNAi mixed with either 10% empty vector (EV) or 10% act‐1. Hits were defined as those that exhibit an observable phenotype when combined with act‐1 RNAi, but not with EV. Scale bar is 500 μm. (c) LifeAct::mRuby is expressed specifically in the body wall muscle cells with the myo‐3 promoter. LifeAct::mRuby binds to F‐actin filaments to allow visualization of actin. Representative fluorescent images of body wall muscle actin are shown. Animals were grown on RNAi from hatch. Animals were heat‐shocked for 1 h at 37°C and immediately imaged. Images were captured on a Zeiss AxioObserver.Z1. Scale bar is 10 μm.