Plasmid Constructs for IRF5 and TRIM21 Studies

Plasmids encoding Myc-tagged IRF5 isoforms were a kind gift of Dr. Frank Neipel (Virologisches Institut - Klinische und Molekulare Virologie, Erlangen, Germany). Plasmids encoding Xpress-TRIM21 and GST-TRIM21 PRY/SPRY domain were a gift from Dr. David Rhodes (Cambridge Institute for Medical Research, Cambridge, UK). HA-ubiquitin wild type and mutants were a gift from Dr. James Burrows (Centre for Cancer Research and Cell Biology, Belfast, UK). Plasmids encoding FLAG-tagged IRF5 full length and deletion mutants were described previously [24] (link). Myc-MyD88 construct was a kind gift from Dr. Alberto Mantovani (Istituto Clinico Humanitas, Milan, Italy). pGL3-IFNA4 luciferase and pGL4-TK-Renilla were a kind gift from Dr. John Hiscott (Lady Davis Institute, Montreal, Canada) and Dr. Kate Fitzgerald (UMASS, Massachusetts, USA), respectively. Plasmids encoding shRNA targeting TRIM21 and scrambled negative control were described previously [16] (link). TLR ligands were purchased from InvivoGen (California, USA). Primary antibodies used were anti-FLAG (Sigma), anti-c-Myc and anti-β-Actin (Abcam), anti-GST (GE Healthcare), anti-Xpress (Invitrogen), anti-IRF5 (Cell Signaling) and anti-α-actinin, anti-HA and anti-TRIM21 (Santa Cruz).