Journal: bioRxiv
Article Title: Enhanced release of ciliary extracellular vesicles suppresses cell migration and promotes cell aggregation
doi: 10.1101/2025.11.30.691462
Figure Lengend Snippet: (A) Immunostaining for primary cilia (red, ARL13B) and nuclei (blue, DAPI) was performed in clone #70 following treatment with siRNAs targeting negative control, IFT88, or KIF3A. Immunofluorescence staining confirmed a marked reduction in the number of ciliated cells after IFT88 or KIF3A knockdown compared with negative control (siNC). Scale bars, 50 µm. (B) Western blots showing ARL13B, IFT88, KIF3A, and GAPDH protein levels in clone #70 after treatment with NC, IFT88, or KIF3A siRNAs. Western blot analysis also demonstrated a significant decrease in IFT88 and KIF3A protein expression by IFT88 and KIF3A knockdown, respectively, relative to siNC. (C) Schematic diagram of the experimental procedure for siRNA knockdown and subsequent EV collection. (D) Western blots of ARL13B, ALIX, and GAPDH in cell lysates and EV pellets collected from clone #70 cultures following treatment with NC, IFT88, or KIF3A siRNAs. Unexpectedly, siRNA-mediated suppression of IFT88 or KIF3A expression increased the amounts of GAPDH, ARL13B, and ALIX in EV pellets compared with siNC-treated controls.
Article Snippet: The following primary antibodies were used for immunofluorescence (IF) and/or western blotting (WB): ARL13B (rabbit polyclonal, 1:1,000 for IF and WB; 17711-1-AP, Proteintech), ARL13B (mouse monoclonal, 1:50 for IF; 66739-1-Ig, Proteintech), FOP (mouse monoclonal, 1:10,000 for IF; H00011116-M01, Abnova), ALIX (rabbit polyclonal, 1:1500 for WB; 12422-1-AP, Proteintech), GAPDH (mouse monoclonal, 1:3,000 for WB; MAB374, Millipore), KIF3A (rabbit polyclonal, 1:1000 for WB; 13930-1-AP, Proteintech), IFT88 (rabbit polyclonal, 1:1,500 for WB; 13967-1-AP, Proteintech).
Techniques: Immunostaining, Negative Control, Immunofluorescence, Staining, Knockdown, Western Blot, Expressing
Proteintech is a verified supplier 
