Expression of pUbq-Cnn-C^T, which ectopically binds γ-TuRCs, reduces the ability of flies to generate progeny. (A) Diagram of normal Cnn-C and chimeric Cnn-C^T in which the CAI domain of Cnn-C (dark blue) is replaced by the shorter N terminus of Cnn-T (red). (B) Graph showing the proportion of embryos that hatched from crosses of WT flies to 0–1- or 1–2-wk-old pUbq-Cnn-C or pUbq-Cnn-C^T males or females, as indicated. Means and 95% CIs are indicated. Total numbers of embryos counted and number of counts are indicated below. (C) Western blots of protein extracts from embryos and testes of WT, pUbq-Cnn-C, and pUbq-Cnn-C^T flies, as indicated. Blots were probed with anti–γ-tubulin, anti–Cnn-C (N-term), anti–Cnn-C (C-term), and anti–Cnn-T^N antibodies as indicated. Note that endogenous Cnn-C (black arrowheads) runs at the same height as pUbq-Cnn-C (blue arrowheads) on these blots, explaining the increased brightness of these bands in the pUbq-Cnn-C extract lanes. Note also that the C-terminal Cnn-C antibody recognizes an unspecific band (asterisks) of approximately the same size as pUbq-Cnn-C^T (red arrowheads) and thus the pUbq-Cnn-C^T band intensity would be lower in the absence of this unspecific band. (D) Western blot showing co-IP of γ-tubulin via anti-Cnn antibodies from embryo extracts expressing either pUbq-Cnn-C or pUbq-Cnn-C^T, as indicated. Red arrowhead indicates Cnn-C^T. Note that, given the low expression of pUbq-Cnn-C^T within embryos, gel loading of the IP lanes was adjusted to better balance the amount of Cnn protein per lane.