<i>sca-1</i> is a target gene of <i>mir-233</i>.

<p>(A) Complementarity between the 3′ UTR of <i>sca-1</i> gene and <i>mir-233</i>. <i>mir-233</i> is presented in the form of DNA from 3′ to 5′ end. WT seeds are marked in red, mutated seeds in blue, Watson-Crick base pairing with a straight line and U-G wobbles with a dotted line. For constructing of <i>sca-1</i>-3′UTR (mut) reporter, the putative <i>mir-233</i> binding site was replaced with an oligonucleotide containing the exact complementary sequence of <i>mir-233</i>. (B) The protein levels of SCA-1 were determined by Western blotting at 8 h after PA14 infection. The blot is typical of three independent experiments. <i>P</i><0.05, <i>mir-233(+)</i> (wild type worms)+PA14 versus <i>mir-233(+)</i>+OP50; <i>P</i><0.01, <i>mir-233(-)</i> (<i>mir-233(n4761)</i> mutant worms)+PA14 versus <i>mir-233(+)</i>+PA14; <i>mir-233(+)</i>+OP50+<i>sca-1</i> RNAi versus <i>mir-233(+)</i>+OP50. (C) Deletion of <i>mir-233</i> did not affect the mRNA levels of <i>sca-1</i>. (D) Fluorescence images of the <i>sca-1</i>-3′ UTR GFP reporter in worms grown on <i>E. coli</i> OP50 or <i>P. aeruginosa</i> PA14. Mutagenesis of the putative binding site for <i>mir-233</i> in the <i>sca-1</i>-3′UTR led to derepression of the GFP expression in <i>mir-233(+)</i> worms after PA14 infection. Meanwhile, the GFP expression was significantly higher in <i>mir-233(−)</i> worms than that in <i>mir-233(+)</i> worms after PA14 infection. The lower panel shows quantification of GFP/mCherry ratio. The data are expressed as percent of control (the ‘wt’ construct on OP50). ^a<i>P</i><0.01 versus <i>mir-233(+)</i>+wt on OP50; ^b<i>P</i><0.01 versus <i>mir-233(+)</i>+wt on PA14.