Figure 2.

Hierarchically recruited PcGs are essential for the transcription of Sens. (A-B) Immunostaining of wing discs expressing attp40-Pc^WT (A, A') or attp40-Pc^Δ69-70 (B, B') driven by AG4 using anti-Sens antibody (red) and anti-Pc antibody (green). Overexpression of Pc^WT has no obvious effect on the level of Sens while Pc^Δ69-70 downregulates the Sens level in the clones marked by dashed lines (Pc positive). (C) ChIP-qPCR analysis using control IgG (grey) or anti-H3K27me3 antibody (black) with WT wing discs (Act5c-Gal4). ChIP signal levels are represented as percentages of input chromatin. RPII140 and Ubx serve as negative and positive controls, respectively. (D-F) RT-qPCR analysis of mRNA levels of E(z) (D), Sens (E) and Ubx (F) in Act5c-Gal4 or Act5c-Gal4/uas-dsRNA E(z) wing discs. (G-J') Wing discs carrying E(z)⁷³¹ or Su(z)12⁴ mutant clones were immunostained with anti-Sens antibody (red) or anti-H3K27me3 antibody (blue); GFP negative clones are marked by dashed lines. Means ± SD; n = 3. ^**P < 0.01. Unpaired, two-tailed Student's t-test. See also Supplementary information, Figure S2.