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. Author manuscript; available in PMC: 2015 Oct 16.
Published in final edited form as: Cell Rep. 2015 Oct 1;13(2):315–325. doi: 10.1016/j.celrep.2015.09.001

Figure 4. Sda is required for accumulation of mature DE-Cadherin to maintain stem cells and hub cells in the Drosophila testicular niche.

Figure 4

(A) Quantification of GSCs, Zfh-1-positive cells and hub cells in testes from E-Cad, N-Cad/+, or sda/+, or E-Cad, N-Cad/+; sda/+ males. P-value between any two genotypes < 0.01. (B-C) Testes from upd-Gal4; sda (B) and upd-Gal4; UAS-E-Cad; sda (C) males stained with anti-Zfh-1, anti-Vasa and anti-Arm; dots indicate GSCs. Asterisk: hub area. Scale bar: 10μm. (D) Quantification of GSCs, Zfh-1-positive cells and hub cells in the testicular niche from upd-Gal4, or upd-Gal4; sda, or upd-Gal4; UAS-E-Cad; sda, or upd-Gal4; UAS-N-Cad; sda D15 males. For GSCs: P-value between any two genotypes < 0.01; for Zfh-1-positive cells and hub cells: P-value between any two genotypes except upd-Gal4; sda is n.s.; whereas P-value between any other genotype and upd-Gal4; sda < 0.01. (E) Quantification of GSCs, Zfh-1-positive cells and hub cells in the testicular niche from tj-Gal4, or tj-Gal4; sda, or tj-Gal4; UAS-E-Cad; sda, or tj-Gal4; UAS-N-Cad; sda D15 males. For GSCs: P-value between any two genotypes < 0.01; for Zfh-1-positive cells and hub cells: P-value between any two genotypes except tj-Gal4; sda is n.s.; whereas P-value between any other genotype and tj-Gal4; sda < 0.01. (F) Quantification of GSCs, Zfh-1-positive cells and hub cells in the testicular niche from nos-Gal4 or nos-Gal4; sda or nos-Gal4; UAS-E-Cad; sda or nos-Gal4; UAS-N-Cad; sda D15 males. For GSCs, Zfh-1-positive cells and hub cells: P-value between any two genotypes except nos-Gal4 is n.s.; whereas P-value between any other genotype and nos-Gal4 < 0.01. (G) Quantitative RT-PCR to measure E-Cad transcript levels in WT and sda mutant testes based on three independent experiments. (H) Immunoblot to measure mature E-Cad protein (~150 KD) levels in WT and sda mutant testes, the 200 KD unprocessed E-Cad is undetectable; CP190 (~190KD) is used as a loading control. (I) Quantification of GSCs, Zfh-1-positive cells and hub cells in testes from sda, or WT, or CadNM12 (N-Cadunprocessable/+), or nos-Gal4; dNc (nos>E-Cadunprocessable), or upd-Gal4; dNc (upd>E-Cadunprocessable) males. For GSCs, P-value between any two genotypes except WT and N-Cadunprocessable/+ is n.s.; P-value between any other genotype and WT < 0.01, and P-value between any other genotype and N-Cadunprocessable/+ < 0.05. For Zfh-1-positive cells: P-value between any two genotypes except WT and upd>E-Cadunprocessable is n.s.; P-value between any other genotype and WT or upd>E-Cadunprocessable < 0.01. For hub cells, P-value between any two genotypes except WT and N-Cadunprocessable/+ is n.s.; P-value between any other genotype and WT or Cadunprocessable/+ < 0.05. Error bar: 95% CI of SEM in (A), (D-F) and (I), P-value calculated by one-tailed t-test.