Cd107a lamp 1

The following monoclonal antibodies (mAbs) were used, conjugated with fluorescein isothiocyanate (FITC), phycoerythrin (PE), or peridinin chlorophyll protein complex (PerCP): anti-CD11c, anti-H2K^b, anti-GR1, anti-CD8, the lysosomal membrane proteins CD107a (LAMP-1) (BD Pharmingen), and the OVA 257-264 (SIINFEKL) peptide bound to H2K^b antibody (25D1.16; eBiosciences). To perform staining, cells were incubated with the corresponding conjugated antibody, or isotype-matched control antibody, for 20 min. Finally, cells were washed once again and analyzed by flow cytometry. Data were collected using a FACScalibur flow cytometer and analyzed with CellQuest software (Becton Dickinson, San Jose, CA).

The antibody clones used for flow cytometry were CD8 (BioLegend, 53-6.7), CD62L (eBioscience, MEL-14), CD44 (eBioscience, IM7) CD107a/LAMP1 (BD Biosciences, H4A3) and NKG7 (Cell Signaling Technology, E6S2A); for microscopy were EEA1 (Cell Signaling Technology, E9Q6G), Rab27 (Cell Signaling Technology, E907E), CD107a/LAMP1 (BioLegend, H4A3) Granzyme B (BD Biosciences, GB11) and rabbit and mouse anti-tubulin (Rockland). Secondary antibodies conjugated to Alexa Fluorophores, ProLong™ Gold Antifade Mountant with DAPI, CellTrace™ Violet and CFSE dyes were purchased from Molecular Probes (Thermo Fisher). Calcein-AM and LysoTracker™ were purchased from Invitrogen (Thermo Fisher).