(A) Confirmation of knockdown of endogenous S100A8 and S100A9 in TR146 cells
using PCR analysis. (B) MMP2 mRNA levels in TR146,
TR146-shRNA-control, TR146-S100A8/A9-shRNA cell lines determined using
quantitative RT-PCR. All mRNA values were normalized to GAPDH,
a housekeeping gene, and expressed relative to TR146 control cells. (C) Levels
of MMP-2 protein in TR146, TR146-shRNA-control, and TR146-S100A8/A9-shRNA cells
as determined using Western blots. Protein levels were normalized to
β-actin and expressed relative to TR146 cells. Assays were performed in
triplicate and repeated three times. The protein and mRNA levels were analyzed
by one-way ANOVA and statistical differences between the cell lines were
reported (P < 0.05).