Fig. S4

Tissue-specific promoter constructs drive expression in a tissue restricted manner; restoring nlrc3-like expression using the myeloid pu.1 driver rescues microglia in nlrc3-like^–/– mutants, Related to Figure 5.

(A) Top row, merged images showing lyz:EGFP stable transgene expression that marks neutrophils and control mCherry expression under the control of different tissue promoters as indicated at 2 dpf. Neutrophil reporter lyz:EGFP provides a reference to indicate the locations where leukocytes normally reside, and to distinguish the neutrophil lyz promoter from the macrophage mpeg1 driver. Top row, merged images of mCherry and EGFP expression; middle row, mCherry expression only from the construct injected; bottom row, stable neurtrophil transgene lyz:EGFP expression only. (B) Schematic showing the Gal4/UAS system applied to drive nlrc3-like in early myeloid cells under the control of the pu.1 promoter. Bar graph showing that 41% of mutants having both the pu.1: Gal4- UAS-EGFP driver and UAS-nlrc3-like construct are rescued, with > 5 microglia. There was no rescue in mutants missing at least one component of the Gal4/UAS system for nlrc3-like expression. (C) Representative images showing rescue of a 4 dpf mutant using the Gal4/UAS system (right). Top row, neutral red staining to visualize microglia, and bottom row, images showing UAS-GFP expression in the same larvae as above, indicating robust activity of the pu.1:Gal4 driver. Number below bar graph represents n, number of mutants analyzed.