Figure 4.

Lid physically interacts with dMyc. (A) Western blot of wild-type (lane 1), GMM (lane 2), and GMM heterozygous for lid¹⁰⁴²⁴ (lane 3) using 24 eye discs per lane. Top panel was probed with anti-dMyc; bottom panel shows anti-Histone H3 loading control. (B) Anti-Lid (left panel) and anti-dMyc (right panel) immunoprecipitations and Western blots for dMyc, Lid, Brm, Osa, and Ash2. A high-molecular-weight Ash2 is detected in both anti-Lid and anti-dMyc immunoprecipitates (arrow). (*) A nonspecific band. (C) Lid interacts with the C-terminal region of dMyc in vitro. In vitro assay testing binding of ³⁵S-methionine Lid to GST (lane 2), GST-dMycN (lane 3), GST-dMycC (lane 4), or GST-dMax (lane 5). (D) In vitro binding assay using ³⁵S-methionine-labeled Rbp-2 and GST (lane 2), c-mycN (lane 3), and c-mycbHLHZ (lane 4). (E) Real-time PCR analysis of dmyc, lid, and Nop60B expression levels relative to expression in GMR/+ controls in eye discs from GMM or GMM discs heterozygous for lid¹⁰⁴²⁴. Asterisk indicates that Nop60B levels in GMM and GMM heterozygous for lid are significantly different (Student’s t-test; P = 0.003).