Fig 2. Knockdown of dBRWD3 suppresses ectopic expression of Ubx in Pc or E(z) depleted brains and condensation failure of Pc or E(z) depleted ventral nerve cords.
(A) A schematic diagram of a 3rd instar central nervous system. The black, red and blue squares indicate the regions examined for the central nervous system, central brain, and ventral nerve cord, respectively. (B and C) Ubx protein levels in the ventral nerve cord (B) and brains (C) of flies expressing mCD8-GFP under the control of Elav-GAL4. The average number of Ubx positive neurons per brain is 0, n = 15. The average length of the ventral nerve cord is 395.6μm, n = 21. (D-F) Ubx protein levels in the ventral nerve cords (D and E) and brains (D and F) of flies expressing UAS-Pc-dsRNA, UAS-mCD8-GFP under the control of Elav-GAL4 to deplete Pc expression. The average number of Ubx positive neurons per brain is 97.1, n = 15, p<0.00001 vs. GAL4 control by Student's t-test. The average length of ventral nerve cord is 468.5μm, n = 49, p<0.001 vs. GAL4 control by Student's t-test. (G and H) Ubx protein levels in the ventral nerve cord (G) and brains (H) of Pc, dBRWD3 doubly depleted flies. The average Ubx positive neurons per brain is 1.9, n = 15, p<0.0001 vs. knockdown of Pc by Student's t-test. The average length of ventral nerve cords is 389.4μm, n = 45, p<0.00001 vs. knockdown of Pc by Student's t-test. (I) Ubx protein (arrowhead) levels in the ventral nerve cord of dBRWD3 depleted flies. (J-L) Ubx protein levels in the ventral nerve cords (J and K) and brains (J and L) of flies expressing UAS-E(z)-dsRNA and UAS-mCD8-GFP by Elav-GAL4. The average number of Ubx positive neurons per brain is 321.6, n = 10, p<0.0001 vs. GAL4 control by Student's t-test. The average length of ventral nerve cords is 456.5μm, n = 39, p<0.01 vs. GAL4 control by Student's t-test. (M and N) Ubx protein levels in the ventral nerve cord (M) and brains (N) of E(z), dBRWD3 doubly depleted flies. The average number of Ubx positive neurons per brain is 69.9, n = 10, p<0.0001 vs. knockdown of E(z) by Student's t-test. The average length of the ventral nerve cords is 393.9μm, n = 40, p<0.001 vs. knockdown of E(z) by Student's t-test. (O and P) Suppression of Pc (O) or E(z) (P) depletion-induced ectopic expression of Antp by dBRWD3 depletion. The number of Antp positive neurons in Pc, dBRWD3 (O), E(z), dBRWD3 (P) doubly depleted brains. *** indicates p<0.0001 by Student's t-test. Brackets indicate the length of the ventral nerve cord. Scale bars indicate 100μm in B, D, G, I, J, and M and 50μm in C, E, F, H, K, L and N.