Figure 6

YPFS suppresses DDP-induced NF-κB activation in A549/DDP cells.

(A) Cultured A549/DDP cells were treated with or without YPFS (1 mg/mL) for 12 hours, followed by 48 hours of DDP treatment at various concentrations. The amounts of phosphor-IKKβ (~87 kDa), IKKβ (~87 kDa), IKKα (~85 kDa) and NF-κB p65 subunit (~65 kDa) were determined by using specific antibodies (left panel). α-Tubulin (~55 kDa) served as a control. The band intensity was calibrated (right panel). Values are in fold of change (X Basal) to control (no drug treatment). (B) Immunofluorescence staining localization of NF-κB p65 by antibody and nuclei staining by DAPI, in A549/DDP cells. Treatment was as that in (A), except 20 μM DDP was used. Asterisks indicate nuclei with NF-κB staining, also the enlarged cells in bottom right corner (left panel). Co-localization coefficients, co-localizing pixel for NF-κB p65 in channel 1 (Ch1) were calculated relative to the total number of pixels for the nuclei (T1) by using Zeiss co-localization coefficient function software (right panel). Bar = 20 μM. Results are expressed as the mean ± SEM from three separate experiments. n = 3. *p < 0.05, **p < 0.01, or ***p < 0.001.