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. 2017 Jul 3;216(7):2131–2150. doi: 10.1083/jcb.201611138

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© 2017 Monis et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

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Figure 2. — Fibrocystin, polycystin-2, and smoothened trafficking rates to the primary cilium. (A–C) Linear regression slope value analysis of newly synthesized CD8-FibrocystinCTS-SNAP, Polycystin-2-GFP-SNAP, or Smoothened-SNAP-GFP trafficking from the Golgi apparatus to the primary cilium when IFT20, GMAP210 (D–F), Exo70 (G–I), Sec8 (J–L), pallidin (M–O), or dysbindin (P–R) is knocked down using lenti-shRNAs. Linear regression analysis of newly synthesized membrane protein delivery to the cilium was performed on selected time points: 0, 1, and 2 h for CD8-fibrocystinCTS-SNAP; 0, 1, 2, 4, and 6 h for polycystin-2-GFP-SNAP; and 0, 1, 2, 4, and 6 h for smoothened-SNAP-GFP. The time points were chosen by determining which data points remained linear before (or if) reaching a plateau in the control groups (Fig. 1; and Figs. S2, S3, and S4). The slope values obtained from this analysis represent the trafficking rates of newly synthesized membrane protein delivery to the cilium. Slope values between control and experimental shRNA groups were compared with one another to determine statistical significance. shRNAs of interest are listed vertically on the left side, and their corresponding membrane proteins are listed horizontally. Error bars represent SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. The IFT20 shRNA linear regression analysis (A–C) from Fig. 1 is shown again for completion.