Recombinant ccl19

Transwell inserts with an 8 micron pore-size (VWR) were placed on 24-well plates containing 1 μg/ml recombinant CCL19 (Peprotech). Naïve or BCG-stimulated BMDCs (3 x 10⁵) were added to the transwell inserts and incubated at 37°C, 5% CO₂. Transwells were removed 2hrs later and the number of DCs that reached the bottom of the plate were enumerated by Trypan blue exclusion.

Astaxanthin (mol wt 596.84), LPS derived from Escherichia coli 026: B6, FITC-Dextran (mol wt 40,000) and Cobalt protoporphyrin (CoPP, a HO-1 inducer) were from Sigma-Aldrich. Alexa Fluor 647-Dextran (mol wt 10,000) was from Thermo Fisher. Carboxyfluorescein succinimidylester (CFSE) and RPMI 1640 medium were from Invitrogen. Fetal bovine serum (FBS) was from Hyclone. Recombinant CCL19, GM-CSF, and IL-4 were from Peprotech. CCK-8 kit was from Beyotime. CD4⁺ T cell isolation kit was from Miltenyi Biotech. Fluorescent-labeled anti-mouse mAbs, PerCP-Cy5.5 CD69, FITC-MHCII, PE-CD40, PE-CD80, FITC-CD86, PE-CCR7 or respective isotype controls, were from BD PharMingen. Alexa Fluor 647 HO-1 or respective isotype was from Abcam. PE-Nrf2 or respective isotype was from Cell Signaling Technology. Tin protoporphyrin IX (SnPP, a HO-1 inhibitor) was from MedChemExpress.