pMIGR1-mP52 and pMIGR1-mGM-CSF were generated by inserting mouse p52 (encoding amino acids 1–405 of NF-κB2) and mouse Csf2 cDNAs, respectively, into the EcoRI and BglII sites of pMIGR1-GFP vector. pcDNA-based expression vectors encoding FLAG-tagged mouse p52 (p52-cFlag-pcDNA3), mouse c-Rel (c-Rel-cFlag-pcDNA3), and mouse RelB (RelB-cFlag-pcDNA3) were from Addgene. The Csf2 promoter luciferase plasmid (pGL3-mCsf2), covering the region −225 to +26 of mouse Csf2 promoter (15 (link)), was provided by Dr. Takeshi Matsumura (Kumamoto University).
Functional grade anti–mouse (m) CD3ɛ (145-2C11) and anti-mCD28 (37.51) antibodies and blocking antibodies for mIFN-γ (XMG1.2) and mIL-4 (11B11) were from eBioscience. Fluorescence-labeled antibodies for mCD4 (L3T4), mCD8 (53-6.7), mCD3 (145-2C11), CD44 (IM7), CD62L (MEL-14), IL-17A (eBio17B7), GM-CSF (MP1-22E9), and IFN-γ (XMG1.2) were purchased from eBioscience. Antibodies for mouse p50 (C-19), c-Rel (sc-70), RelB (C-19), NIK (H-248), Lamin B (C-20), and Hsp60 (H-1) were from Santa Cruz Biotechnology. An antibody recognizing both p100 and p52 (Anti-p52/p100) was provided by NCI Preclinical Repository.
Functional grade anti–mouse (m) CD3ɛ (145-2C11) and anti-mCD28 (37.51) antibodies and blocking antibodies for mIFN-γ (XMG1.2) and mIL-4 (11B11) were from eBioscience. Fluorescence-labeled antibodies for mCD4 (L3T4), mCD8 (53-6.7), mCD3 (145-2C11), CD44 (IM7), CD62L (MEL-14), IL-17A (eBio17B7), GM-CSF (MP1-22E9), and IFN-γ (XMG1.2) were purchased from eBioscience. Antibodies for mouse p50 (C-19), c-Rel (sc-70), RelB (C-19), NIK (H-248), Lamin B (C-20), and Hsp60 (H-1) were from Santa Cruz Biotechnology. An antibody recognizing both p100 and p52 (Anti-p52/p100) was provided by NCI Preclinical Repository.