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Figure 2 | Arthritis Research & Therapy

Figure 2

From: The metastatic tumor antigen 1-transglutaminase-2 pathway is involved in self-limitation of monosodium urate crystal-induced inflammation by upregulating TGF-β1

Figure 2

Inverse effect of transglutaminase 2 (TG2) knockdown or overexpression on monosodium urate (MSU) crystal-induced cytokines production in RAW264.7 cells. (A) RT-PCR analysis of IL-1β, TNF-α, TG2 and TGF-β1 mRNA expression in RAW264.7 cells transfected first with TG2 specific small interfering RNAs (siRNAs) for 48 h, then stimulated with or without MSU crystals (1 mg/ml) for 4 h. (B) Quantification of RT-PCR analysis of TG2, TGF-β1, IL-1β and TNF-α mRNA expressions. (C) Cells transfected with TG2-siRNA were stimulated with or without MSU crystals (1 mg/ml) for 36 h. The levels of TGF-β1, IL-1β and TNF-α in supernatants were collected and evaluated by ELISA. Mean control value of TGF-β1 (209.5 ± 21.2 pg/ml), IL-1β (23.1 ± 2.1 pg/ml) and TNF-α (137.8 ± 15.2 pg/ml). (D) RT-PCR analysis of IL-1β, TNF-α, TG2 and TGF-β1 mRNA expression in RAW264.7 cells stably transfected with TG2 or empty vector (mock) stimulated with or without MSU crystals (1 mg/ml) for 4 h. (E) Immunoblot analysis of TG2 in RAW264.7 cells stably transfected with TG2 or empty vector (mock). (F-H) Quantification of RT-PCR analysis of (F) TGF-β1, (G) IL-1β and (H) TNF-α mRNA expression. (I) Cells transfected with TG2 or empty vector were stimulated with or without MSU crystals (1 mg/ml) for 36 h. The levels of TGF-β1, IL-1β and TNF-α in supernatants were collected and evaluated by ELISA. The results are expressed as the means ± SEM from three independent experiments (n  3, * = P < 0.05, ** = P < 0.01 compared with control (100%)).

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