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Fig. 3 | Epigenetics & Chromatin

Fig. 3

From: Genome-wide RNAi screen in Drosophila reveals Enok as a novel trithorax group regulator

Fig. 3

Mutants of enok genetically interact with Pc and trx. Two mutants of enok (enok1 and enok2) crossed to either Pc1 (a) or PcXL5 (b) alleles strongly suppressed extra sex comb phenotype. +/Pc heterozygotes from Pc alleles (Pc1 and PcXL5) crossed to w1118 flies represent control and showed strong extra sex combs. More than 150 male flies with the desired genotype (enok mutant/Pc mutant) in the progeny of each cross were analyzed. Based on the number of extra sex comb hairs on 2nd and 3rd legs, male flies were categorized (see key) [62]. c enok1 and enok2 were crossed to trx1 and trxE2 alleles and males in the progeny were scored for loss of pigmentation (A5 to A4 transformation, white asterisk). trx/+ heterozygotes from w1118 crossed to trx mutants served as control. As compared to trx/+ , double mutants (enok; trx) showed strong enhancement of trx mutant phenotype (A5 to A4 transformation). ac Percentage of flies in each category was plotted as bar graphs with SEM. d Effect of enok mutations on the expression of homeotic (abd-A, Abd-B, Ubx, Dfd) and non-homeotic (pnt) genes. qRT-PCR performed on 20-h-old homozygous enok1 or enok2 embryos compared to w1118 embryos of the same age. A drastic reduction was observed in the expression of all the genes analyzed. eh As compared to Ubx expression pattern (PS5-PS12) in w1118 used as a control (e, f), a strongly reduced Ubx expression is seen in stage 15 homozygous enok embryos. Misaligned (arrows) and merged (oval) parasegmental boundaries of Ubx staining patterns are visible (g, h). As compared to w1118 embryos (i, j), immunostaining of enok mutant embryos showed strongly reduced Abd-B expression restricted to PS13-14 only. ad Experiments were performed in triplicates and individual Student’s t tests were performed for each category and the p values obtained were marked as **p ≤ 0.01, ***p ≤ 0.001 or ****p ≤ 0.0001. Error bars represent SEM

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