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Fig. 4 | Cell & Bioscience

Fig. 4

From: 1-Palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol (PLAG) attenuates gemcitabine-induced neutrophil extravasation

Fig. 4

PLAG attenuates gemcitabine-induced activation of NOX2 by inhibiting membrane localization of cytosolic subunits Rac1 and p47phox phosphorylation. a The effect of DPI on gemcitabine-induced CXCL8 mRNA expression was analyzed by RT-PCR. b THP-1 cells were treated with various doses of DPI 1 h before gemcitabine treatment, and the protein level of CXCL8 in the supernatant was determined by ELISA. c The effect of DPI on gemcitabine-induced phosphorylation of ERK, p38 MAPK and JNK was analyzed by western blot in THP-1 cells. d Transmigration of dHL-60 cells towards THP-1 conditioned medium was decreased by DPI treatment. e THP-1 cells were treated with 100 μg/ml of PLAG 1 h before gemcitabine treatment, and then were processed for immunofluorescence with anti-Rac1 antibody. The same cells were also stained with DAPI to visualize nuclei. f THP-1 cells were stimulated with 10 μg/ml of gemcitabine for 0 to 2 h, or g the cells were pretreated with PLAG for 1 h and then stimulated with gemcitabine for 2 h, and the cytosolic and membrane proteins were fractioned as described in “Materials and methods”. The separated proteins were subject to western blot analysis and blotted with antibodies against: Rac1, Na/K-ATPase, and α-tubulin. h Whole lysates of THP-1 cells were separated by SDS-PAGE and subject to western blot. The blot was probed with anti-phospho-p47phox antibody. *p < 0.05, **p < 0.01, ***p < 0.001

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