11. Clamp both H2O tubing (see Note 11).

12. Flow 140 mL of 1% Triton X-100 into the lumen for 30 min

using the syringe pump at flow rate of 4.5 mL/min (Fig. 5).

Used Triton X-100 should end up in the waste bottle.

13. Clamp Triton X-100 tubing (see Note 11).

14. Remove the PBS tubing clamp.

Fig. 4 H2O step schematic representation. SDS, Triton X-100, and PBS tubes are clamped on top of their

respective reservoirs. H2O flows through the tubing main circuitry, to the trachea and ends in the waste bottle

Fig. 5 Triton X-100 step schematic representation. H2O and PBS tubes are clamped on top of their respective

reservoirs. Triton X-100 washes the trachea and ends in the waste bottle. SDS circuitry remains closed until

the end of the de-epithelialization

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Alba E. Marin-Araujo et al.