Regulation of dMyc and Gro shared targets. (A) Venn diagram of dMyc/dMax/dMnt and Gro targets (see SI Text). (B) Functional distribution of the 37 dMyc and Gro shared targets identified by DamID (plus one target identified experimentally; SI Table 2). (C) dMyc is required for expression of dMyc/Gro targets. Western blot analyses indicating that dMyc/Gro target gene expression is reduced in S2 cells treated with dMyc RNAi (dMyc-i). Kr, Krüppel; γTub, γTubulin; vnd, ventral nerve defective; cycB, cyclin B; fibril, fibrillarin. (D and E) dMyc is required for dMyc/Gro target gene transcription. (D) Western blot analysis of dMyc and Gro in untreated S2 cells or in cells treated with either dMyc or control (GFP) RNAi. (E) Northern blot analysis of Nop60B expression using the same extracts depicted in D. (F) Decline in target gene expression because of reduced dMyc levels can be partially restored by the simultaneous reduction of Gro levels. Western blot analysis of S2 cells treated with RNAi to the indicated gene(s) and assayed for the level of dMyc, Gro, or the shared target proteins Nop60B, CycA, CycB, γTub, Barren (Barr), and the direct Myc-Max target, fibrillarin. Relative quantification of the proteins is given below the lanes with the first 0 μg of dMyc-i RNAi lane (wild type) being set to 100 (see Materials and Methods).