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Figure 2: Cigarette smoke condensate could increase neprilysin membrane distribution and block its endocytosis. (a) BEAS-2B cell was culture and treated with cigarette smoke condensate (25 μg/ml) for 96 h and then subjected to biotination and Western blots. (b-d) Quantification for surface neprilysin, internalized neprilysin, and total neprilysin from (a), and normalized with GAPDH (n = 4). (e) BEAS-2B cell with indicated treatment was stained live for surface (green) and internalized (red) neprilysin. After fixation, total neprilysin was also stained (blue). (f) Quantification for internalized/surface neprilysin level from (e) (n = 15). (g) BEAS-2B cell with indicated treatment was lysate for mRNA extraction and followed by a reverse transcript polymerase chain reaction for neprilysin mRNA level. (h) Quantification for neprilysin mRNA level from (g), normalized with GAPDH (n = 3). Data were represent here as mean ± standard error of mean (*P < 0.05, **P < 0.01), scale bar is 10 μm

Figure 2: Cigarette smoke condensate could increase neprilysin membrane distribution and block its endocytosis. (a) BEAS-2B cell was culture and treated with cigarette smoke condensate (25 μg/ml) for 96 h and then subjected to biotination and Western blots. (b-d) Quantification for surface neprilysin, internalized neprilysin, and total neprilysin from (a), and normalized with GAPDH (<i>n</i> = 4). (e) BEAS-2B cell with indicated treatment was stained live for surface (green) and internalized (red) neprilysin. After fixation, total neprilysin was also stained (blue). (f) Quantification for internalized/surface neprilysin level from (e) (<i>n</i> = 15). (g) BEAS-2B cell with indicated treatment was lysate for mRNA extraction and followed by a reverse transcript polymerase chain reaction for neprilysin mRNA level. (h) Quantification for neprilysin mRNA level from (g), normalized with GAPDH (<i>n</i> = 3). Data were represent here as mean ± standard error of mean (*<i>P</i> < 0.05, **<i>P</i> < 0.01), scale bar is 10 μm