Figure 4

Confirmation of the effect of the novel method, time- and cell-specific RNAi (T.C.RNAi). (A) The average recovery ratio of GFP intensity of AWC neurons in animals without RNAi constructs, animals with only gcy-10p::gfp(s), gcy-10p::gfp(as), hsp16-2::gfp(s), hsp16-2::gfp(as), animals with both gcy-10p::gfp(s) and hsp16-2::gfp(as) or both hsp16-2::gfp(s) and gcy-10p::gfp(as) after heat-shocked or control condition. (n ≥9 animals). (B) The average recovery ratio of GFP intensity of AWB and AWC neurons in animals with srd-17p::gfp(s) and hsp16-2::gfp(as) after heat-shocked or control condition (n ≥10 animals). (C) Representative images of gcy-10p::GFP in animals with srd-17p::gfp(s) and hsp16-2::gfp(as) before (left panel, intact) and after (center panel, photobleach) photobleaching and after heat-shocked condition (right panel, heat shock). Arrow heads indicate AWB neurons, and arrows indicate AWC neurons. Scale bars = 10 μm. (D) The average recovery ratio of GFP intensity of anterior or lateral protrusions of vulva in animals without RNAi constructs or animals with myo-3p::gfp(s) and hsp16-2::gfp(as) after heat-shocked or control condition (n ≥14 protrusions of vulva). (E) Chemotaxis to isoamyl alcohol (1:1000) in wild-type animals, odr-3 mutants and wild-type animals with odr-3 T.C.RNAi (n ≥4 assays). AWC::(s) and hsp::(as) mean gcy-10p::odr-3(s) and hsp16-2::odr-3(as). Error bars represent SEM and asterisks indicate significant differences (*P <0.05, **P <0.01, Dunnett’s test or Student’s t-test). SEM, standard error of the mean.