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. 2018 Jul 6;50(7):1–9. doi: 10.1038/s12276-018-0114-1

Fig. 3. CSE, but not NE, upregulates PAR2 expression.

Fig. 3

a BEAS-2B cells were treated with CSE (0.5 and 1%) for the indicated times. The expression of PAR2 was measured by quantitative real-time PCR. Data were normalized to the expression of GAPDH. Data represent the mean ± SD; **P < 0.05. b Cells were stimulated with CSE (1%) for 24 h. The cells were fixed and permeabilized for 10 min. Immunofluorescent staining of PAR2 was performed using an anti-PAR2 antibody, followed by an Alexa Fluor 488 antibody. Cells were analyzed using an ECLIPSE TE300 (Nikon) fluorescence microscope. The mean intensity of PAR2 per cell number was determined using ImageJ. c, d BEAS-2B cells were treated with CSE (0.5, 1, and 2%) or NE (1 U/ml) for the indicated times. Total cell lysates were extracted and subjected to western blot analysis for PAR2 and GAPDH. e, f Cells were treated with CSE (0.5, 1, 2, and 4%) or NE (1 U/ml) for 24 h. The membrane fraction was isolated and then subjected to western blot analysis for PAR2 using an antibody to detect the N-terminal extracellular domain of PAR2. Results are representative of three independent experiments