Abd-B and EcR signaling drive Beat-Va _L cell death but not Beat-Va _M remodeling. (A–C) Beat-Va neurons genetically labeled with mCD8::GFP and driving a UAS-Abd-B^RNAi at wL3 (A), 2 h APF (B), and 6 h APF (C) stained for cleaved Dcp-1. Magnified views of A3–A7 lateral cells from the right and left are shown as a single plane image on the right of each image. (D) Beat-Va neurons labeled with mCD8::GFP and driving UAS-Abd-B^RNAi at wL3 stained with the EcR-B1 showing the continued colocalization of EcR-B1 in the absence of Abd-B. (E) Beat-Va neurons at wL3 genetically labeled with mCD8::GFP and crossed to UAS-Abd-B and stained with anti-Abd-B. The two most anterior lateral and medial cell bodies are magnified and show Abd-B expression, which is not usually present in these cells, confirming overexpression was successful. (F) The three most anterior medial Beat-Va neurons at wL3 labeled with CD8::GFP and stained with Abd-B shows Abd-B is present in some medial cells even though they do not undergo cell death. (G) Beat-Va_M neurons at HE genetically labeled with mCD8::GFP and stained with anti-Abd-B. The A5–7 medial cells continue to express Abd-B at HE. Left and right hemisegment denoted by “L” and “R.” (H) Beat-Va neurons at wL3 genetically labeled CD8::GFP (green) stained with the EcR-B1 Receptor (magenta) showing all lateral and medial cell bodies express the receptor. (I) Beat-Va neurons at 6 h APF genetically labeled with CD8::GFP (green), expressing a UAS regulated EcR^DN, and stained with the EcR-B1 Receptor (magenta) to identify cells that express UAS-EcR^DN. There are typically no EcR-B1 positive cells at this time. (J) Beat-Va neurons at wL3 genetically labeled CD8::GFP (green) and expressing UAS-Cas9 and EcR-targeting gRNA ubiquitously. Stained with the EcR-B1 Receptor (magenta) to evaluate if cells lose EcR-B1 expression. (K) Quantification of gRNA against usp, plum, EcR on Beat-Va EcR-B1 expression when compared with control. One-way ANOVA, Kruskal–Wallis test for multiple comparisons. n = number of animals. Colored data points correspond to the representative image shown in the figure. All error bars are SEM. Scale bar is 5 µm in all images with a single cell body and 20 µm in all other images.