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. 2021 Feb 17;7(8):eabe2597. doi: 10.1126/sciadv.abe2597

Fig. 4. ISWI knockdown during separable developmental windows leads to distinct behavioral phenotypes.

Fig. 4

Quantification of rhythmicity as measured by maximum FFT amplitude of temporally restricted ISWI knockdown (blue) through (A) first, (B) second, and (C) mid-third instars (left to right) compared to constitutive knockdown (red) and genetic controls (black and grays) [for all elav > UAS-ISWI RNAi (red), n = 23; from left to right for all other conditions: knockdown through first instar, n = 25, 28, 27, and 27; knockdown through second instar, n = 30, 32, 30, and 30; knockdown through mid-third instar, n = 29, 31, 30, and18]. (D) Representative images of FasII immunostaining with ISWI knockdown through first (top) and second instars (bottom), with quantification of percentage of brains with normal MB morphology (for knockdown through first instar: n = 12, black; n = 15, blue; for knockdown through second instar: n = 21, black; n = 22, blue) (Fisher’s exact test). Scale bars, 50 μm. (E) Quantification of PER assay with temporally restricted ISWI knockdown through second instar (blue, n = 14) compared to genetic controls (black, n = 10; gray, n = 12) (two-way ANOVA with post hoc multiple comparison test; asterisks denote significance of ISWI knockdown condition compared to both genetic controls in post hoc testing). (F and G) Courtship index (left) and copulation success (right) for ISWI knockdown through F) mid-third (n = 39, 62, and 42 from left to right) and (G) early pupation (n = 60, 81, and 49 from left to right). (H) Summary of windows of ISWI knockdown that give rise to different adult phenotypes.