PMC

Plasma device characteristics. (A) Schematic diagram of the plasma device and experimental setup. The cells were cultured on glass cover slips for plasma treatment. (B) The air plasma source is shown.

Cellular expression of p-FAK, and uptake of GNP and p-FAK-GNP. (A) Western blot shows the expression of p-FAK protein on HaCaT and G361 (p <0.001). (B) The uptake of GNP is shown. The green spots represent GNP absorbed by G361 cells. (C) The uptake of p-FAK-GNP is shown. The p-FAK-GNP specifically bind to p-FAK and generate fluorescence around the G361 cell membranes.

The impact of inducing of p-FAK-GNP with plasma on selective and immediate cell death in G361 cells. (A) After plasma treatment, the cell death rate was measured using Trypan blue assay and compared to the untreated controls (G361: p-FAK-GNP and plasma, p < 0.01). (B) In untreated G361 cells (left panel), cells treated with only plasma (middle panel), and cells treated with p-FAK-GNP and plasma (right panel) (2500 ×). (C) After incubation, the G361 cells were subjected analysis by western blot using NEU, p-NEU, FAK, p-FAK, and p-paxillin antibodies as indicated.

Effects of anti p-FAK and p-FAK-GNP on the proliferation of G361 and HaCaT cells. (A and B) After 24 h incubation, cell viability was measured using a WST-1 assay. The G361 cells were treated with anti-p-FAK antibodies and p-FAK-GNP for the indicated time periods, and the viability was compared to untreated cells (p-FAK-GNP 24 h, p <0.05; p-FAK-GNP 48 h and 72 h, p < 0.01; anti-p-FAK 48 h and 72 h, p < 0.05). (C) After 24 h incubation, the cell viability was measured using a WST-1 assay. The HaCaT cells were treated with p-FAK-GNP for the indicated time periods. (D) After incubating the G361 cells with p-FAK-GNP for the indicated times, the cells were harvested and lysates were analyzed by western blot using p-paxillin, p-FAK, p-NEU, FAK, and NEU antibodies as indicated.

Induction of apoptosis in G361 cells by p-FAK-GNP. (A) The G361 cells were treated with p-FAK-GNP for 12 to 48 h. At each time point, the cells were subjected to FACS analysis using PI solution as indicated. (B) After a 24 h incubation in the presence (lower panel) or absence (upper panel) of p-FAK-GNP, the translocation of cytochrome c was visualized by immunocytochemistry using an anti-cytochrome c antibody. (C) After 24 h incubation in the presence (lower panel) or absence (upper panel) of p-FAK-GNPs, the subcellular localization of AIF was visualized by immunocytochemistry using an anti-AIF antibody. (D) The G361 cells were incubated in the presence of p-FAK-GNP for the indicated times. The total protein lysates were used for SDS-PAGE followed by western blots using antibodies against BAX, Bdl-2, Caspase 3, Caspase 9, PARP-1, DFF 45, and GAPDH (as a loading control).