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Response to Comment on “An extracellular matrix fragment drives epithelial remodeling and airway hyperresponsiveness”

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Science Translational Medicine  19 Jun 2019:
Vol. 11, Issue 497, eaaw0462
DOI: 10.1126/scitranslmed.aaw0462

Figures

  • Fig. 1 PGP is comparable to AcPGP at inducing proliferation of human bronchial epithelial cells.

    Undifferentiated normal human bronchial epithelial cells (Lonza) were cultured in media or media supplemented with media alone, AcPGP (10 ng/ml), or PGP (10 or 100 ng/ml) and visualized over a period of 72 hours using a JuLI Stage automated cell imaging system. (A) Fold change in cell confluence over a 72-hour period depicted for individual wells in each treatment group. (B) After 72 hours, the cells were fixed and stained for Ki-67, with Ki-67–positive cells expressed as a percentage of 4′,6-diamidino-2-phenylindole–positive cells. Figure represents data combined from two independent experiments with five wells per group in each experiment. Results are depicted as means ± SEM. **P < 0.01, ***P < 0.001, and ****P < 0.0001 using analysis of variance (ANOVA) with Bonferroni correction.

  • Fig. 2 AcPGP induces IL-8 and G-CSF release from human bronchial epithelial cells.

    (A and C) Normal human bronchial epithelial cells were cultured at air-liquid interface. Respective wells were treated with media or media supplemented with AcPGP (10 μg/ml). Apical supernatants were collected on day 1 for measurement of IL-8 (A) and G-CSF (C) by enzyme-linked immunosorbent assay (ELISA). (B and D) Undifferentiated normal human bronchial epithelial cells were cultured in media or media supplemented with AcPGP (10 ng/ml) or PGP (100 ng/ml) for 72 hours. In some groups, AcPGP-treated cells were preincubated with either anti-CXCR1/2 antibodies or IgG2a isotype control antibody. Supernatant was collected for the measurement of IL-8 (B) and G-CSF (D) by ELISA. (A) and (C) present combined data from two independent experiments with two wells per group in each experiment. (B) and (D) represent data combined from two independent experiments with five wells per group in each experiment. Results are depicted as means ± SEM. *P < 0.05, **P < 0.01, and ****P < 0.0001, using Mann-Whitney statistical test (A and C) or ANOVA with Bonferroni correction (B and D).

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