PMC

TLR signaling proteins in T1DM. A, NFKβ activity in control (C) and T1DM monocytes was performed using TransAM reagents as described in Subjects and Methods. *, P < 0.001 compared with controls. B, Representative Western blotting results of TLR downstream signaling proteins MyD88, pIRAK-1, and Trif was performed using specific rabbit antibodies to the respective (phospho)proteins as described in Subjects and Methods using β-actin as loading and internal control for MyD88 and Trif and IRAK for pIRAK-1. C, Control. Densitometric ratios. *, P < 0.05 vs. control. Controls and T1DM, n = 31 per group.

TLR2 and TLR4 expression in T1DM. Surface expression of TLR2 (resting and Pam3CSK4 activated) (A) and surface expression of TLR4 (resting and LPS activated) (B) on human monocytes isolated from control and T1DM patients (n = 31 per group) were assessed by flow cytometry as described in Subjects and Methods. Data are expressed as mean fluorescence intensity units (mfi). *, P < 0.05 vs. control basal; #, P < 0.05 vs. control activated. C, TLR2 and TLR4 mRNA in T1DM. Representative RT-PCR results for TLR2 and TLR mRNA from three different controls and three T1DM patients were assessed as described in Subjects and Methods using GAPDH as control. Lower panel, The TLR2 and TLR4 to GAPDH ratio, respectively, for all control and T1DM patients (n = 31 per group). *, P < 0.05 vs. control.