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bab gene functions contribute to regulation of Engrailed/Invected accumulation in Terminal Filament cells but not in Cap Cells.

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posted on 2020-11-05, 18:28 authored by Laurine Miscopein Saler, Virginie Hauser, Mathieu Bartoletti, Charlotte Mallart, Marianne Malartre, Laura Lebrun, Anne-Marie Pret, Laurent Théodore, Fabienne Chalvet, Sophie Netter

(A-C') Prepupal ovaries immunostained for detection of GFP (green) and Engrailed/Invected (En/Inv) (grey in A’ and red in B’,C’). F-actin labeling is shown in red (A) and grey (B-C'). Anterior is up, medial is left. Scale bars: 10 μm. (A-A', B') Green and yellow brackets indicate Terminal Filament (TF) cells and Cap Cells (CCs), respectively. (C-C') The cluster of medial hhG+ cells depleted of Bab1 and Bab2 is encircled (pink dotted line). Green and yellow arrowheads indicate the anterior- and posterior-most hhG+ cells, respectively. (A-A’) Niche region of a mosaic ovary (hs-FLP; FRT-babAR07/FRT-GFP) containing mitotic cell clones homozygous for the babAR07 mutation that are marked by absence of GFP (green and yellow arrowheads). In babAR07 mutant TF cells (green arrowheads), the signal for En/Inv is lower than that in wild type TF cells (green brackets). However, in babAR07 mutant Cap Cells (CCs) in contact with GCs (yellow arrowheads), the level of En/Inv is similar to its endogenous level in wild type CCs (yellow bracket). (B,C) Prepupal ovaries expressing GFP in hhG+ cells (hhG>GFP), and in C, RNAi transgenes against bab1 and bab2 (hhG>GFP, bab1IR, bab2IR) as well. (B’,C’) Higher magnifications of the regions framed with dotted lines in (B,C). The anterior-most cells hhG+ cells of hhG>GFP, bab1IR, bab2IR ovaries (C’, green arrowheads) present lower levels of En/Inv (C’, green arrowheads) than TF cells in the control (B’, green brackets). The posterior-most hhG+ cells depleted of Bab1 and Bab2 (C', yellow arrowheads) show a similar level of En/Inv than in control CCs (B’, yellow brackets). (D) Graph comparing the fluorescence intensity (arbitrary units) of En/Inv in control niche and babAR07 clonal cells. In babAR07 mutant TF cells (hs-FLP, babAR07), the En/Inv fluorescence intensity is more than 2-fold lower than in adjacent control TF cells (hs-FLP, GFP). However, in babAR07 mutant CCs, the En/Inv fluorescence intensity is similar to that in adjacent control CCs. (E) Graph comparing the fluorescence intensity (arbitrary units) of En/Inv in CCs in the control (B', yellow brackets) and in posterior-most hhG+ cells knocked down for bab1 and bab2 using RNAi (C', yellow arrowheads). Values are presented as means +s.d. p-values are calculated using a Kruskal-Wallis (D) or Mann-Whitney (E) test. n: sample size; NS: Not Significant (p>0.05); **** p<0.0001).