Mouse anti gt335

Transfected cells were fixed in 100% methanol at −20°C for 10 min. After washing with 1× PBS, the cells were blocked with blocking solution (4% donkey serum in PBST) at RT for 1 h. Cells were incubated with primary antibodies: mouse anti‐acetylated α tubulin (Sigma, T7451, MO, USA, 1:2,000), mouse anti‐GT335 (Adipogen, AG‐20B‐0020, CA, USA, 1:2,000), rabbit anti‐γ tubulin (Santa Cruz, SC‐10732, CA, USA 1:200), rabbit anti‐ARL13b (Proteintech, 17711‐1‐AP, IL, USA, 1:1,000) antibodies at 4°C overnight in blocking solution. After washing with 1× PBS, the cells were incubated with Alexa Flour^® 488‐conjugated secondary antibodies (Invitrogen, mouse A11001, rabbit A11008, 1:500) or Alexa Flour^® 594‐conjugated secondary antibodies (Invitrogen, mouse A11005, rabbit A11012, 1:500) at RT for 1 h. Randomly selected fields (≥ 5) were photographed using the confocal microscope (Carl Zeiss, LSM 700 or LSM 780, Germany) in all of imaging analyses. The intensity and length of cilia were measured using ImageJ software (NIH, USA).