Commercial antibodies used were as follows: anti-Acetylated tubulin (
Actub, clone 6-11B-1, 1/10000,
T6793, Sigma), anti-Gamma-tubulin (GTU-88, 1/1000,
T6557, Sigma), anti-b-actin (clone AC-15, 1/30000, A5441, Sigma), anti-PACSIN1 (1/100, 196 003, Synaptic Systems), anti-PACSIN2 (1/250,
ab37615, Abcam), anti-PACSIN2 (1/500, 10518-2-AP, Proteintech), anti-PACSIN3 (1/100,
ab37612, Abcam), anti-Pericentrin (PCTN, 1/5000,
NB100-61071, Novus Biologicals), anti-EHD1 (EPR4954, 1/500, ab109311, Novus Biologicals), anti-RPGRIP1L (1/200, 55160-1-AP, Proteintech), anti-TMEM67 (1/200, 12780-1-AP, Proteintech), anti-CEP164 (1/500,
sc-240226, Santa Cruz), anti-CP110 (1/1000, 12780-1-AP, Proteintech), anti-CEP97 (1/1000, A301-945A, Bethyl), anti-Arl13b (1/300, clone N295B/66, NeuroMab/UC Davis), anti-GFP Alexa 488 (1/1000,
A21311, Molecular Probes Life Technologies), Phalloidin conjugated with Alexa 488 (1/50,
A12379, Molecular Probes Life Technologies),
Hoechst (1/3000,
H3570, Molecular Probes Life Technologies) and all secondary antibodies were from Life Technologies. The rabbit anti-RAB8A antibody was a gift from Johan Peränen (University Helsinki, Finland). SNAP-Cell647-SiR reagent was purchased from New England Biolabs. Nocodazole (Calbiochem) was from Sigma. Doxycyclin hydrochloride was obtained from Sigma and used according to manufacturer’s instruction.
Insinna C., Lu Q., Teixeira I., Harned A., Semler E.M., Stauffer J., Magidson V., Tiwari A., Kenworthy A.K., Narayan K, & Westlake C.J. (2019). Investigation of F-BAR domain PACSIN proteins uncovers membrane tubulation function in cilia assembly and transport. Nature Communications, 10, 428.