HEK-BlueTM hTLR4 cells (InvivoGen, San Diego, CA, USA) were plated at a density of 2.5 × 104 cells/well with Ps-K18 in HEK-Blue detection media (InvivoGen, San Diego, CA, USA). After 1 h, the cells were stimulated by 20 ng/mL LPS and incubated for 8 h at 37 °C in a humidified 5% CO2 atmosphere. SEAP activity was determined by measuring absorbance at 630 nm.
RT-PCR was performed to measure inhibition activity of Ps-K18 on expression of TLR4 mediating inflammatory response signaling as previously reported [55 (link)]. The results visualized by UV illumination using Alpha Innotech gel documentation system (Alphaimager ® HP; Alpha Innotech Corporation, San Leandro, CA, USA).
RT-PCR was performed to measure inhibition activity of Ps-K18 on expression of TLR4 mediating inflammatory response signaling as previously reported [55 (link)]. The results visualized by UV illumination using Alpha Innotech gel documentation system (Alphaimager ® HP; Alpha Innotech Corporation, San Leandro, CA, USA).
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