Anti lamp1 clone d2d11

Recombinant human M-CSF was purchased from Miltenyi Biotec and recombinant human IFN-γ from Becton Dickinson. M-CSF was used at a concentration of 50 ng/ml and IFN-γ at a concentration of 10 ng/ml. Dexamethasone, hydrocortisone, imatinib mesylate, and mifepristone were from Sigma-Aldrich and used at the following concentrations: dexamethasone 10–1000 nM, hydrocortisone 300 nM, imatinib mesylate 5–10 µM, and mifepristone 1 µM. 25D3 and 1,25D3 were obtained from Biomol and used as indicated. Bafilomycin A1 was purchased from Sigma-Aldrich and Invivogen and used at 100 nM. The mAbs used were: anti–IL-15 PE (clone 34559) and IgG1 PE isotype control (clone 11711) from R&D Systems; anti-LAMP1 (clone D2D11) from Cell Signaling; anti-LAMP1 (clone eBioH4A3) from eBioscience; IgG1 pure isotype control (clone X40) and IgG2a isotype control (clone X39) from Becton Dickinson; anti-cathelicidin (clone OSX12) from Abcam or Novus Biologicals; and monoclonal anti-TCIRG1 from Abcam. Anti-mouse IgG1 PE (clone A85-1) and anti-mouse IgG2a FITC (clone R19-15) were obtained from Becton Dickinson. LysoSensor Green DND-189, DAPI, and secondary Abs labeled with Alexa Fluor 488 or Alexa Fluor 594 were purchased from Life Technologies. FCS and human AB serum were purchased from PAA Laboratories and PAN Biotech. RPMI 1640 and macrophage serum-free media (SFM) were obtained from Life Technologies.