Fig. 3: CCK expression in ASM and in the lung. | Nature Communications

Fig. 3: CCK expression in ASM and in the lung.

From: Antagonizing cholecystokinin A receptor in the lung attenuates obesity-induced airway hyperresponsiveness

Fig. 3

a qRT measurement of CCK mRNA expression in airway cells. FIB: lung fibroblasts; HBE: human bronchial epithelia cells; HBE(D0): day 0 of initiation of air-liquid interface (ALI) culture; HBE(D14): day 14 post-ALI; ASM: airway smooth muscle cells. (n = 3 biologically independent samples per group). b Immunostaining of mouse lung tissue section for CCK (green). α-SMA (α-smooth muscle actin, red) was used as a marker of ASM. Scale bar = 25 µm. Shown is a representative image from three independent experiments with similar results. c qRT measurement of CCK mRNA expression in ASM cells after treatment with indicated free fatty acids (250 μM) or control (isopropanol). n = 4 biologically independent samples per group; hexanoic vs. control (P = 0.0772); 9-decenoic vs. control (P < 0.0001); dodecanoic vs. control (P = 0.0533); palmitoleic vs. control (P < 0.0001); myristoleic vs. control (P = 0.0007). Data represent the mean ± SEM; two-tailed, unpaired t-test (*P < 0.05). d ELISA measurement of CCK in ASM cell culture medium after free fatty acid treatment (n = 4 biologically independent samples per group); palmitoleic acid vs. control (P = 0.0019); myristoleic acid vs. control (P = 0.0280). Data represent the mean ± SEM; one-tailed, unpaired t-test (*P < 0.05). e qRT measurement of CCK mRNA expression in the lungs of HFD-fed mice vs. regular chow-fed mice. n = 6 mice per group. Data represent the mean ± SEM; P = 0.0132; one-tailed, unpaired t-test (*P < 0.05). f CCK protein expression in the lungs of HFD-fed mice vs. regular chow-fed mice. n = 4 mice per group. Data represent the mean ± SEM; P < 0.0001; one-tailed, unpaired t-test (*P < 0.05). g qRT measurement of CCK mRNA expression in the airways (lungs and trachea) db/db mice compared to wild type control (WT). n = 4 mice per group; Data represent the mean ± SEM; P = 0.0152; one-tailed, unpaired t-test (*P < 0.05). h CCK protein expression in the lungs of db/db mice (n = 4) vs. wild-type control (WT). n = 4 mice per group; Data represent the mean ± SEM; P = 0.0353; one-tailed, unpaired t-test (*P < 0.05). Actin or 18 s rRNA was used as reference gene. Source data are provided as a Source Data file.

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