Fig. 1: Expression of a rare-codon-enriched reporter during differentiation of Drosophila neurons.

A Schematic of GFP- rare^b and GFP-common reporters. 5’ is to the left. B Expression of GFP-rare^b in whole WL3 larvae, scale bar is 1 mm. Arrow labels the brain. B’ Expression of GFP-rare^b in one lobe of a WL3 brain, scale bar is 20 µm. C-C’ Expression of GFP-common in WL3 larvae as in B, B’. D Schematic of neuroblast lineage in Drosophila larval brain, showing markers of neuroblasts (NBs), ganglion mother cells (GMCs) and neurons in magenta. E–J Representative images of WL3 brain expressing GFP (green), either GFP-rare^b (E, G, I) or GFP-common (F, H, J). Brains are co-labeled with cell type markers (magenta). Deadpan (Dpn) marks NBs (E, F), earmuff (erm) GAL4 drives membrane-targeted tdTomato in GMCs (G, H), and Elav marks neurons (I, J). Solid outline shows cells expressing marker and GFP protein. Scale bar is 10 μm. E, G, I GFP-rare^b reporter, dashed outline shows cells expressing marker that do not express GFP protein. K-L’: Representative images of WL3 brains expressing GFP-rare^b (green on left, white on right) co-labeled with Dpn (magenta on left) to mark NBs. Scale bar is 20 μm. K-K’ GFP-rare^b reporter brain. L-L’ GFP-rare^b reporter brain with inscuteable (insc) GAL4 driving UAS-brat RNAi in the NB lineage. M Quantification of area of central brain covered by GFP-rare^b expressing cells in areas affected by each RNAi. Each data point = 1 animal, mean and SEM plotted. Two biological replicates conducted, n for each genotype are as follows: no RNAi = 7, brat RNAi = 5, numb RNAi = 5, pros RNAi = 5. All comparisons by one-way ANOVA followed by Dunnett’s multiple comparisons tests comparing no RNAi to: brat RNAi, p = 0.0008; numb RNAi, p = 0.0378, pros RNAi, p = 0.0412. Source data are provided as a Source Data file.