Extended Data Fig. 3: Cert are required for migrasome formation.
a, L929 cells expressing TSPAN4–GFP were infected with control (NS) or two Cert-shRNA lentiviruses and stained with mCherry–NT-Lys. Green, TSPAN4–GFP; red, mCherry–NT-Lys. Scale bar, 10 µm. b,c, qPCR analysis of Cert expression in cells from Fig. 4e and Extended Data Fig. 3a. Data are the mean ± s.d., n = 3 independent experiments. ****P < 0.0001; two-tailed unpaired Student’s t-test. d, Statistical analysis of migrasome numbers from Extended Data Fig. 3a. n = 150 cells per group from three independent experiments. Data are presented as the median with the interquartile range. ****P < 0.0001 (NS versus Cert-KD-2; NS versus Cert-KD-3); two-tailed unpaired non-parametric test (Mann–Whitney test). e, L929 cells stably expressing TSPAN4–GFP were infected with lentivirus carrying non-specific-shRNA (NS) or Cert-shRNA (Cert-KD-1) constructs. Cells were stained with anti-ceramide (α-Ceramide) and observed by TIRF. Green, TSPAN4–GFP; red, anti-ceramide. Scale bars, 10 µm. f, The number of ceramide foci in RFs per cell from Extended Data Fig. 3e was statistically analysed. NS, n = 180 cells, Cert-KD-1, n = 157 cells. Data (from three independent experiments) are presented as the median with the interquartile range. ****P < 0.0001; two-tailed unpaired non-parametric test (Mann–Whitney test). P < 0.05 was considered statistically significant.
