Figure 7 : MANF suppresses the expressions of NF-κB target genes.

From: Mesencephalic Astrocyte-Derived Neurotrophic Factor Is Involved in Inflammation by Negatively Regulating the NF-κB Pathway

Figure 7

(a–b) MANF knockdown increases the mRNA levels of IL-8 (a) and A20 (b). 293T cells transfected with MANF-siRNA were stimulated with TNF-α (10 ng/ml) for 30 min 24 hrs after transfection. Relative mRNA levels were analyzed by real-time qPCR. The data are expressed as the means ± SD of at least three independent experiments. ** P = 0.0079, # P = 0.0390, compared with the control. (c–d) Recombinant MANF inhibits the mRNA expressions of IL-1β (c) and TNF-α (d) in FLS. The FLS isolated from adjuvant arthritis (AA) rats were cultured in DMEM containing 10 μg/ml LPS and MANF protein for 24 hrs. BSA and DXM were used as the negative control and positive control, respectively. The mRNA levels were determined by RT-PCR. The data are expressed as the means ± SD of at least three independent experiments. In panel c, * P = 0.015, ** P = 0.002, *** P = 0.000, compared with BSA. In panel d, * P = 0.025, ** P = 0.001, *** P = 0.00, compared with BSA. (e–f) MANF-siRNA increases the expressional levels of IL-1β (e) and TNF-α (f) in FLS. The transfected FLS were treated with LPS (10 μg/ml) for 24 hrs. Relative mRNA levels were analyzed by RT-PCR. The data are the means ± SD of at least three independent experiments. ** P = 0.06, ## P = 0.01, compared with siRNA control. (g) MANF-siRNA increases the secretion of IL-1β and TNF-α in FLS. The transfected FLS were treated with LPS (10 μg/ml) for 24 hrs. The levels of IL-1β and TNF-α were analyzed by ELISA. The data are expressed as the means ± SD from at least three independent experiments. * P = 0.036; # P = 0.017, compared with siRNA control. (h) Recombinant MANF decreases the secretion of IL-1β and TNF-α in FLS. The FLS of AA rats were incubated with 10 μg/ml LPS and MANF for 24 hrs. The levels of IL-1β and TNF-α in the medium were determined by ELISA. The data are expressed as the means ± SD from at least three independent experiments. * P = 0.018, ** P = 0.008, *** P = 0.000, # P = 0.03, ## P = 0.019, ### P = 0.001, compared with BSA.