Fig. 3
Effect of HM on TLR4 expression in THP-1 cells. a Representative histograms showing isotype control antibody FITC-conjugated IgG2b (A.1) used to establish settings for the fluorescence-activated cell sorting (FACS) instrument, untreated TLR4-positive THP-1 cells (A.2), and untreated TLR4-negative HEK293 cells (A.3). Bar graph (A.4) shows the percentage of TLR4-positive cells (determined by FACS analysis) in HM-treated THP-1 cells, compared with untreated cells (control). b Representative immunoblot (B.1) showing the effect of various concentrations (15.6, 62.5, and 500 μg/mL) of HM on TLR4 protein expression in THP-1 cells after 24 h of incubation, compared with untreated cells (control). Bar graph (B.2) shows the relative protein expression levels of TLR4, calculated as a ratio of GAPDH expression (the loading control). c Bar graph shows the relative expression levels of TLR4 mRNA, calculated as a ratio of the expression of the house-keeping gene β-actin. Results are presented as the mean ± SD of three independent experiments. (*) and (**) signify a statistically significant difference (p < 0.05 and p < 0.01) compared with the control