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. 2011 Jun 16;7(6):e1002109. doi: 10.1371/journal.pgen.1002109

Figure 2. Testing hypersensitive strains and validating the screen with visible phenotypes.

Figure 2

A: Table indicating the quantification of the laterality defects observed following RNAi knock-down of the indicated genes in wild-type (otIs114) and 3 different RNAi sensitized backgrounds. nre-1(-) lin-15b(-) and eri-1(-); lin-15b are the most sensitive strains for ASE. In all cases n>40. 1Scored with otIs114; 2Scored with otIs186; 3Scored with otIs299; 4ASE defect refers to loss or dimming of ASEL::gfp, ASER::gfp or ASE::gfp; 5ASE defect refers to ectopic expression of ASEL::gfp in ASER or loss or dimming of ASER::gfp or ASE::gfp; 6ASE defect refers to loss or dimming of ASEL::gfp or ASE::gfp and ectopic expression of ASER::gfp in ASEL; 7ASE defect refers to loss or strong dimming of ASEL::gfp, ASER::gfp or ASE::gfp; 8For fozi-1 ASE defect refers to ectopic expression of ASEL::gfp in ASER or loss of ASER::gfp or ASE::gfp. For lim-6 it refers to loss of ASEL::gfp expression or ectopic expression of ASER::gfp in ASEL; 9ASEL::gfp when still present appeared much dimmer than in any other strain; 106/6 escapers displayed ectopic expression of ASEL::gfp in ASER; 111/1 escaper displayed ectopic expression of ASEL::gfp in ASER; 12Taken from Flowers and Poole et al. 2010. B: Representative pictures of the ASE laterality that result from RNAi knock-down of the indicated known ASE factors in an nre-1(-) lin-15b(-) background. ASEL is labeled with lim-6prom::gfp and ASER is labeled with gcy-5prom::gfp. L4440 refers to feeding with an empty RNAi vector. Examples of the 4 main classes of ASE phenotype as described in [18]. Class I is 2-ASEL, class II is 2-ASER, class III is 0-ASE (loss of ASE cell fate) and class IV is mixed (in the case of fozi-1 ASEL markers are depressed in ASER but ASER markers are unaffected). C: Comparison of the visible phenotypes observed in our screen with previous screens for all genes on Chr I. Ovals represent the amount of bacterial clones that gave an RNAi phenotype in an experiment with results from our screen shaded in pink and previous published results shaded in grey. Areas that overlap represent clones (in %) for which in both experiments an RNAi phenotype was detected. Differences and overlap between an RNAi experiment done with the lin-15b nre-1 mutant strain and (in the following order): (1) all data previously reported for an RNAi experiment on Chr. I (whether as part of a genome wide screen, subscreen or a single experiment, exported from wormbase.org WS190); (2) the data obtained by screen done with the hypersensitive rrf-3 strain [22]; (3) the data obtained by screen done with the standard laboratory strain Bristol N2 [23]; (4) the data obtained by an overlap of phenotypes reported in both Simmer and Kamath screens.