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40

60 80

20

SPAN4

0.0

0.5

1.0

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2.0

20% 40% 60% 80%

Folds

ofincreasingof

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igrasomes

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A

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Supplementary information, Figure S1Cell confluence affects the formation of PLSs.

A. TSPAN4-GFP-expressing NRK cells were seeded by serial dilution, cultured for 12

hours to reach 20%, 40%, 60% and 80% confluence, then observed by confocal

microscopy. Scale bar, 10 m. B. Cells from A were assessed for number of PLS per

cell. n=100 cells from three independent experiments. Error bars indicate the s.d.