Extended Data Fig. 5: Tspan4 and Tspan5 enhance cell survival following membrane damage.
From: Assembly of Tetraspanin-enriched macrodomains contains membrane damage to facilitate repair
a Cell death was analysed by PI permeability in NRK Tspan4-OE, NRK WT and NRK Tspan4-KO cells after detergent treatment. Cells were imaged by confocal microscopy. Scale bar, 10 µm. b Statistical analysis of absolute PI intensity in NRK Tspan4-OE, NRK WT and NRK Tspan4-KO cells after detergent treatment. Mean±s.e.m., n = 29 cells for Tspan4-OE, 42 cells for WT and 33 cells for Tspan4-KO from 3 independent experiments. c Verification of the Tspan4-KO Tspan5 knockout cell line by PCR. d Tspan4-OE (Tspan4-overexpression), WT, and Tspan4/Tspan5-KO cells pre-stained by FM 1-43 (5 μM) were subjected to photodamage treatment and the FM 1-43 signal was monitored. Time-lapse images were collected by confocal microscopy and displayed as heatmap images. Scale bar, 10 µm. e Statistical analysis of FM 1-43 fluorescence intensity of the total cell substrate during and after photodamage treatment. Mean±s.e.m. n = 19 cells for Tspan4-OE, 30 cells for WT, 32 cells for Tspan4/Tspan5-KO, 17 cells for Tspan4-OE no-damage, 21 cells for WT no-damage and 9 cells for Tspan4/Tspan5-KO no-damage from 3 independent experiments. Statistical analysis of cell viability in each group (NRK Tspan4-OE, NRK WT, NRK Tspan4-KO, NRK Tspan4/Tspan5 double-knockout cells) after PBS or LPS electroporation treatment. Mean±s.e.m., n = 4 independent experiments. Unpaired t-test was used.
