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. 2012 May 8;10(5):e1001326. doi: 10.1371/journal.pbio.1001326

Figure 7. Elevation of NOX in Ras transformed cells and in primary pancreatic cancer tissues.

Figure 7

(A) Increase of NOX activity in T-Rex 293 Tet/on cells with 1 mo K-rasG12V induction compared with control. Error bars, ±SD. ** p<0.01 (n = 3). (B) NOX activity was substantially elevated in pancreatic K-rasG12V stably transformed HPDE-kRasG12V cells compared with the parental HPDE (human pancreatic ductal epithelial) cells. Error bars, ±SD. * p<0.05 (n = 3). (C) Increased protein level of p22phox in HPDE-K-rasG12V cells and primary pancreatic cancer cells Aspc1 and Panc-1 compared to HPDE cells. (D) Increased NOX activity in the H-RASV12-transformed (T72Ras) cells compared to normal ovarian epithelial cells (T72). Error bars, ±SD. ** p<0.01 (n = 3). (E) Increased gene expression of NOX components (NOX1, NOX2, NOXA1, p22phox, and p47phox) in the H-RASV12-transformed T72Ras cells when compared to the parental T72 cells. Expression of mRNA was measured by qRT-PCR analysis. Error bars, ±SD. * p<0.05; ** p<0.01 (n = 3). (F) Preferential disruption of mitochondrial transmembrane potential by DPI (3–10 µM, 20 h) in H-RASV12-transformed T72Ras cells compared with the parental T72 cells. Mitochondrial transmembrane potential was measured by flow cytometry using rhodamine-123 as a probe. (G) Representative tissue staining showing no expression of p22phox protein in normal pancreas (a, single arrow, normal pancreatic duct; double arrows, islet cells) and chronic pancreatitis (b, arrows, benign pancreatic ducts), and a moderately differentiated pancreatic ductal carcinoma (c) and strong positive staining in a moderately differentiated pancreatic ductal carcinoma (d). The strong positive staining in the inflammatory cells served as internal positive controls for our immunohistochemical stain (original magnification, 200×). Expression of p22phox in stage II pancreatic ductal carcinoma (PDC) and benign pancreatic tissue on microarray. p22phox expression is considered to be significantly different between PDC and benign group and higher in PDC group (p<0.0001 analyzed by Fisher's exact test).