Figure 3
Figure 3. TLR2 increases ROS production and adhesion to extracellular matrix proteins. (A) Meg-01 cells were stained with 10μM 5-(and-6)-carboxy-2′,7′-difluorodihydrofluorescein diacetate, and pretreated for 1 hour with 10μM N-acetyl-L-cysteine or diphenyliodonium. Cells were then treated with 1 μg/mL of Pam3CSK4 (PAM) and analyzed by flow cytometry (n = 6). **P < .01 compared with NT. (B) Meg-01 cells were treated for 3 hours with 1 μg/mL of PAM. Cells were then allowed to adhere to fibronectin-coated coverslips for 1 hour. Adherent cells were stained with FITC-conjugated anti-human CD41 antibody. Cells were counted in 5 separate 20× views. Representative photographs were taken of NT or PAM. (C) Meg-01 cells were treated with 25 μg/mL of IgG or TLR2 antibodies for 1 hour, treated with PAM as before, and allowed to adhere to fibrinogen-coated wells for 1 hour. Adherent cells were measured using MTT in serum free media. Levels were compared with NT (n = 8). **P < .01 and ***P < .001 compared with NT.

TLR2 increases ROS production and adhesion to extracellular matrix proteins. (A) Meg-01 cells were stained with 10μM 5-(and-6)-carboxy-2′,7′-difluorodihydrofluorescein diacetate, and pretreated for 1 hour with 10μM N-acetyl-L-cysteine or diphenyliodonium. Cells were then treated with 1 μg/mL of Pam3CSK4 (PAM) and analyzed by flow cytometry (n = 6). **P < .01 compared with NT. (B) Meg-01 cells were treated for 3 hours with 1 μg/mL of PAM. Cells were then allowed to adhere to fibronectin-coated coverslips for 1 hour. Adherent cells were stained with FITC-conjugated anti-human CD41 antibody. Cells were counted in 5 separate 20× views. Representative photographs were taken of NT or PAM. (C) Meg-01 cells were treated with 25 μg/mL of IgG or TLR2 antibodies for 1 hour, treated with PAM as before, and allowed to adhere to fibrinogen-coated wells for 1 hour. Adherent cells were measured using MTT in serum free media. Levels were compared with NT (n = 8). **P < .01 and ***P < .001 compared with NT.

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