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. Author manuscript; available in PMC: 2023 May 1.
Published in final edited form as: J Thromb Haemost. 2022 Feb 27;20(5):1182–1192. doi: 10.1111/jth.15670

Figure 2: Enhanced Tnfa expression in BMDMs is driven by TG2-cross-linked fibrinogen and not independent effects of TG2.

Figure 2:

(A) BMDMs were cultured on a fibrinogen- or TG2-cross-linked fibrinogen-coated surface or uncoated surface. Immediately after plating, the TG2-specific inhibitor ERW1041E (20 μM) or vehicle (0.1% DMSO, final concentration) was added. Tnfa expression was measured via qRT-PCR after four hours. (B) Fibrinogen was incubated with TG2 or TG2 + 0.1, 1.0, or 2.5 mM of the transglutaminase inhibitor, cystamine. Cross-linked products were resolved on a 4–12% Bis-tris gel and stained with SimplyBlue total protein stain. Lane 1: Molecular weight marker, Lane 2: fibrinogen, Lane 3: empty, Lanes 4–6: Fibrinogen +TG2 with the indicated concentration of cystamine. (C) BMDMs were cultured on unmodified fibrinogen, fibrinogen incubated with 2.5 mM cystamine, TG2-cross-linked fibrinogen, or fibrinogen incubated with TG2 + 2.5 mM cystamine for 4 hours. Tnfa expression was measured via qRT-PCR. Data are presented as mean fold change + SEM compared to BMDMs on uncoated surface (n=3). *p<0.05. (D) BMDMs were cultured on uncoated plates and treated with either unmodified fibrinogen or TG2-cross-linked fibrinogen (10 μg/mL) or vehicle for 4 hours. Tnfa and Il10 expression were measured via qRT-PCR. Data are presented as mean fold change + SEM compared to vehicle-treated BMDMs (n=3).