Figure EV4. siRNA‐based secondary screen to validate substrates.

• A
  Experimental scheme for siRNA screen for hCDC14A substrate(s) essential for cilia length control.
• B–E
  Altogether we analyzed 24 proteins in this secondary screen. 14 proteins were chosen because they were hits in all four experiments (two replicates each for overexpression and BioID MS). Ten proteins appeared in at least two experiments (one replicate each for overexpression and BioID). Indicated proteins were depleted by siRNA during ciliogenesis in RPE1 WT and hCDC14A ^PD cells. Cilia length was subsequently measured by indirect immunofluorescence microscopy. The experiment was repeated twice, and 70–150 cilia were measured for each experiment; Mean ± SD. *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001. One‐way ANOVA. All the other depletions did not significantly affect cilia length compared to the NSC control. Red dotted lines indicate the mean values of NSC.