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. Author manuscript; available in PMC: 2018 Jul 24.
Published in final edited form as: Dev Cell. 2017 Jul 24;42(2):130–142.e7. doi: 10.1016/j.devcel.2017.06.022

Figure 3. The conserved GCL domain is essential for substrate recognition.

Figure 3

(A) Lysates prepared from 0–2 hours AEL (after egg laying) embryos expressing either empty vector (EV) or a FLAG-HA-tagged GCL variant (WT, E100K, Δ40aa, or R377A) were immunoprecipitated (IP) with anti-FLAG resin. Immunocomplexes were probed with antibodies specific to the indicated endogenous proteins. GCL transgenes, generated with UAS promoter and the k10 3′UTR regulatory sequence, were expressed using the germline-specific driver nanos-GAL4∷VP16.

(B) Densitometric scanning quantification of the Torso expression levels in the embryo lysates relative to the corresponding αTubulin (loading control) levels. The averaged relative Torso expression levels from triplicate experiments are plotted with respect to the EV control. Error bars represent standard deviation.

(C) EV or a FLAG-HA-tagged GCL variant (WT, E100K, Δ40aa, R377A) was expressed in gclΔ/Δ embryos using the germline-specific driver nanos-GAL4∷VP16. Number of PGCs in each embryo was counted and plotted. Bars represent the mean ± standard deviation. (n = 20 for each genotype, ****P < 0.0001, ns = not significant, Mann-Whitney test).

See also Figure S3.