Extended Data Fig. 1: Validation of the VPS13C^mClover-Flp-In cell line under the control of tetracycline and effect of the cathepsin C inhibitor E64D on VPS13C recruitment to endolysosomes.
a, Schematic drawing showing the experimental system used for the inducible stable expression of VPS13C^mClover. b, Western blots for the indicated proteins of whole cell lysates from VPS13C^mClover-Flp-In cells under the control of tetracycline, with or without tetracycline (0.1 μg/ml) treatment for 24 hours. Tubulin was used as a loading control. The experiment was repeated three times with similar results. c, Live fluorescence images of VPS13C^mClover-Flp-In cells before and after a 10 min exposure to 1 mM LLOMe with and without the additional presence of the cathepsin inhibitor E64d (200 μM). The experiment was repeated three times with similar results. Quantification of the intensity of the VPS13C^mClover punctate fluorescence per cell from a representative experiment is shown on the right. n = 20 cells (Control or E64D) were analysed. Graph shows the normalized fluorescence relative to time 0. Data were compared using two-sided t-tests. Error bars represent ±SD. **represent P = 0.0029, P = 0.0065 and P = 0.0057, respectively. Numerical data and unprocessed blots are provided.
