Research Paper Volume 13, Issue 4 pp 5804—5823

Figure 9. Knockdown of CD74 affected MIF-induced inflammatory responses in JFs. (A) qRT-PCR analysis of CD44, CXCR4, CXCR7, and CD74 in JFs following treatment with 2 μg/mL recombinant MIF for 24 h. (B) Immunoprecipitation was used to determine the interaction between MIF and CD74. (C) Knockdown efficiency of CD74 siRNAs were tested using qRT-PCR after transfection for 48 h and siRNA2 was chosen for subsequent experiments. (D) Western blot analysis of CD74 following siRNA2 knockdown of CD74 for 48 h. siRNA (control) with the same nucleotide composition as siRNA2 but lacking sequence homology to the CD74 was designed as a negative control. (E) Expression of Tnf-α, Il-1β, and Il-6 was assessed via qRT-PCR following treatment of JFs with siRNA2 or control for 48 h and stimulation with 2 μg/mL recombinant MIF for 24 h. Error bars represent standard deviation. *P <0.05 compared with the control group.