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Fig. 3 | Respiratory Research

Fig. 3

From: Azithromycin induces epidermal differentiation and multivesicular bodies in airway epithelia

Fig. 3

Confirmation of genes and proteins upregulated after Azm treatment. a) Expression of four genes identified in the top 30 list (Table 2). Expression was confirmed using qRT-PCR of RNA isolated from 22-day cultures treated with Azm compared to control. RNA was pooled from duplicate wells and each sample was measured in triplicate in qPCR. Data shown is averaged from 3 separate experiments (*P < 0.05; ***P < 0.001). Abbreviations: CRNN-cornulin; KRT1-cytokeratin 1; SPINK5-serine protease inhibitor Kazal-type 5; DSG1-desmoglein 1. b) Expression of epidermal associated markers in VA10 cells after Azm treatment. ALI cultures from 22 day treatment with Azm were stained using immunohistochemistry for proteins shown to have enhanced gene expression in the sequencing data. Azm-treated samples stained positive for epidermal markers cytokeratin 1, desmoplakin, desmoglein 2 and corneodesmosin, in addition to the tight junction protein claudin 1. In addition to positive staining, notable thickened cell layers were observed after Azm treatment. Cells were counter-stained with hematoxylin. Bar = 100 μm

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