<i>gei-11</i> knockdown effect through regulation of cholinergic receptors at the NMJ.

Abstract

<p>(A) Real-time qPCR analysis of AChR subunits <i>unc-29</i>, <i>unc-38</i>, <i>unc-63</i>, <i>lev-1</i> and <i>acr-16</i>, and GABA_R<i>unc-49</i>, in 6 day old wt animals fed with <i>gei-11</i> RNAi. Data are normalized to the levels of each gene on vector-treated wt animals (±SD). (B) Suppression of Q35 aggregation by <i>gei-11</i> RNAi was abolished by co-treatment with L-AChR (<i>unc-38, unc-63, unc-29</i>) but not with N-AChR (<i>acr-16</i>) subunits RNAi (±SD). Individual RNAi controls are shown in light grey (also see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003711#pgen.1003711.s008" target="_blank">Table S1</a>). (C) Cholinergic sensitivity assay: 5 day old animals treated with <i>gei-11</i> or vector RNAi were scored for paralysis on 1 mM levamisole plates (±SD). L-AChR mutant animals <i>unc-38(e264)</i>, <i>unc-63(x26)</i> and <i>unc-29(e1072)</i> were used as controls. Two-way ANOVA and Bonferroni test ***<i>p</i><0.001 relative to vector control. (D) AChR antagonist dTBC (2.5 mM in water) prevented suppression of Q35 aggregation by <i>gei-11</i> RNAi (±SD). Q35;<i>unc-38(e264)</i> is a control for AChR-dependent effect. Student t-test ***<i>p</i><0.001. (E) Real-time qPCR analysis of AChR subunits <i>unc-29</i>, <i>unc-38</i> and <i>unc-63</i> upon muscle-specific <i>gei-11</i> RNAi (<i>rde-1(ne219);m</i>RDE-1, 6 days old), relative to vector control (±SD). (F) Cholinergic sensitivity assay: 5 day old wt, <i>rde-1(ne219);m</i>RDE-1 and <i>rde-1(ne219)</i> animals treated with <i>gei-11</i> or vector RNAi were scored for paralysis on 1 mM levamisole plates (±SD). Two-way ANOVA and Bonferroni test ***<i>p</i><0.001, **<i>p</i><0.01, *<i>p</i><0.05 relative to vector control. (G) Aggregation quantification upon <i>gei-11</i> RNAi in Q35, Q35;<i>rde-1(ne219);m</i>RDE-1 (muscle-specific RNAi) and Q35;<i>rde-1(ne219)</i> (impaired RNAi); shown as a relative % to Q35;vector (±SD). Student t-test ***<i>p</i><0.001, ns/not significant.</p