TECPR1 conjugates LC3 to damaged endomembranes upon detection of sphingomyelin exposure

A

Schematic of TECPR1 domain structure. AIR, ATG5‐interacting region; PH, Plekstrin homology domain. Tectonin repeat indicated.

B

Confocal micrographs of HeLa cells expressing the indicated GFP‐tagged TECPR1 constructs, infected with S. Typhimurium, fixed at 1 h postinfection and DNA stained with DAPI. Scale bar, 20 μm.

C

Percentage of S. Typhimurium positive for the indicated constructs in HeLa cells, fixed at the indicated timepoints postinfection. Mean ± SEM of 3–4 independent experiments performed in duplicate. n > 100 bacteria per coverslip.

D

Confocal micrographs of HeLa cells expressing TECPR1 N‐terminal (N′) DysF‐GFP infected with mCherry‐expressing S. Typhimurium, fixed at 30 min postinfection. Scale bar, 20 μm.

E

Confocal micrographs of HeLa cells expressing TECPR1 N′ DysF‐GFP with or without FLAG‐nSMase2, infected with S. Typhimurium, fixed at 30 min postinfection and stained with anti‐Galectin 8 antibody. DNA stained with DAPI. Scale bar, 20 μm.

F

Percentage of S. Typhimurium positive for N′ DysF‐GFP in Hela cells, expressing FLAG‐nSMase 2 or not, fixed at 30 min postinfection. Mean ± SD of two independent experiments performed in duplicate.

G, H

Confocal micrographs of HeLa cells expressing the indicated GFP‐tagged DysF domains, infected with mCherry‐S. Typhimurium and fixed at 30 min postinfection (G) or treated with osmotic shock assay, fixed and stained with anti‐Galectin 8 antibody (H). The bottom row in H depicts an enlarged version of the boxed region shown above. Scale bar, 20 μm.

Figure 2. The N‐terminal DysF domain of TECPR1 detects sphingomyelin.