Fig. 2.

Conservation, phenotypes, and methylation pattern for UAS-E(z) variant. a) Domain structure schematic of human EZH1(Hsa_EZH1) and Drosophila melanogaster E(z)(Dmel_E(z)) b-b′) Conservation of EZH1-pA678 across species and also in human paralogs EZH1 and EZH2 (The structures are derived from UniProt) [Hsa_EZH1: Homo Sapiens EZH1; Dis R: Disordered Region; SANT: SANT-Myb- Protein binding Domain, CXC: cysteine-rich DNA binding domain; Dme_E(z): D. melanogaster Enhancer of zeste [E(z)]. c–i) Overexpression assay of UAS-E(z) constructs with different GAL4 lines. The progenies of GAL4 driven UAS-E(z) flies are evaluated for patterning defects, such as A4 segment pigmentation (lateral view), presence of extra sex combs on the second pair of legs (L2) and extra wing veins. Actin-GAL4/UAS-E(z)^WT flies are represented in c–e. The phenotypes observed in Actin-GAL4/UAS-E(z)^A691G flies are represented in f–h. The penetrance of patterning variation seen in male progeny of different ubiquitous or tissue-specific GAL4-driven UAS-E(z) flies is represented in i (n ≥ 20). The extra wing-vein patterning defect is also present in females. j–j′): Western blot assay for H3K27me3 of UAS-E(z)^WT and UAS-E(z)^A691G overexpression with Actin-GAL4 and elav-GAL4. Trimethylation of H3K27 was evaluated in the Actin-GAL4 and elav-GAL4 driven UAS-E(z)^WT and UAS-E(z)^A691G adult progenies (j). j′) is the graphical representation of the H3K27me3 values normalized to H3 values. ActinGAL4 P = 0.0087, elavGAL4 P = 0.1091. UAS-lacZ was used as a positive control. Unpaired t-tests determined P-values.