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Figure 5. LPS-induced TLR4 expression. CHME-5 cells were stimulated with LPS (1 μg/mL) at 37 °C for 3, 6, and 18 h. A: TLR4 mRNA expression was analyzed using RT-PCR analysis with β-actin as housekeeping gene, 3 h (****P < 0.0001) and 6 h (**P < 0.006) vs. US. Image is representative of 3 independent experiments (n = 3) for each treatment group; B: CHME-5 cells were stimulated with LPS (1 μg/mL) at 37 °C for 10-270 min, whole cell lysates were subjected to SDS-PAGE electrophoresis and immunoblotted with TLR4 (1:1,000) and β-tubulin (1:1,000) antibodies; C: integrated density, 90 min (**P < 0.005) and 270 min (**P < 0.01) vs. US. Images are representative of 5 independent experiments (n = 5) for each treatment group. Bars are presented as mean ± SEM. LPS: lipopolysaccharide; TLR4: toll-like receptor 4; RT-PCR: real-time polymerase chain reaction; US: unstimulated; SDS-PAGE: sodium dodecyl sulfate polyacrylamide gel electrophoresis