Pf 429242

UA (Sigma-Aldrich) was dissolved in DMSO and used at a final concentration of 1 µM. AnCoA4 was dissolved in DMSO and used at a final concentration of 50 µM. PF-429242 (Tocris) was dissolved in DMSO and used at a final concentration of 500 nM. TG (Sigma-Aldrich) was dissolved in DMSO and used at a final concentration of 500 nM. Ionomycin (Sigma-Aldrich) was dissolved in DMSO and used at a final concentration of 2.5 to 5 µM. ML-SA1 (Sigma-Aldrich) was dissolved in DMSO and used at a final concentration of 10 µM.

Reagents utilized in the in vitro culture system include human IL-2 (Roche); IFN-γ (Sigma); IL-6, IFN-α and IL-21 (PeproTech); goat anti-human F(ab’)₂ fragments (anti-IgM and -IgG; Jackson Immunoresearch); HybridoMax hybridoma growth supplement (Gentaur); Lipid Mixture 1, chemically defined (200X) and MEM Amino Acids Solution (50X; Sigma); and PF-429242 (Tocris Bioscience).
The antibodies employed for protein detection include CREB3L2 (NBP1-88697, Novus Biologicals), ATF6 (40256; Novus Biological), ACTIN (A1978; Sigma), LC3A/B (Cell Signaling). The reagents utilized in flow cytometry assays include PE-conjugated anti-CD19 (LT19; Miltenyi Biotech), V450-conjugated anti-CD20 (2H7; E-Biosciences), PE-Cy7-conjugated anti-CD38 (HB7; BD Biosciences); APC-conjugated anti-CD138 (B-B4; Miltenyi Biotech); CountBright beads (Invitrogen) and 7-AAD (BD Biosciences).

Reagents used for the in vitro culture system include Ex527 (Cat. #E7034, Sigma-Aldrich®, Deisenhofen, Germany), PF429242 (Cat. #3354, Tocris Bioscience, Bristol, UK), TPh A (Cat. #TPh A ab144376, Abcam®, Cambridge, MA, USA), and Human Pirin ELISA Kit (Cat. #MBS280347, MyBioSource, San Diego CA, USA).