Fig. 2.

Schematic representation of the interval-based secretomics workflow. ① Treatments are performed in the presence of serum and secretion is probed in 2 hour serum-free collection windows. Serum-free supernatants are cleared to remove potential cell debris by centrifugation through a 0.45 μm filter plate. ② Proteins are digested; peptides are chemically labeled with isobaric TMT reagents and are subsequently pooled. ③ Pooled peptide samples are analyzed by LC-MS/MS.