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. 2016 Sep 10;5(3):35. doi: 10.3390/cells5030035

Figure 4.

Figure 4

K8 and K19 are upregulated after the depletion of microbiota by oral antibiotic treatment. Mice (n = 3 for controls, lanes 1–3 in (A), n = 7 for antibiotic-treated, lanes 4–10 in (A); strain FVB/n) were treated with or without oral broad-spectrum antibiotics for 56 days and samples for protein, RNA and immunofluorescence analysis were collected at the end of the treatment. The samples were then analyzed to assess changes in keratin levels in response to the depletion of microbiota. (A,B) The levels of the main colonic keratins K8 and K19 were upregulated after antibiotic-treatment, along with increased levels of stress-responsive HSF2 and K8 pS74 when quantified as in Figure 1 and shown as average ± SD. * p < 0.05, *** p < 0.001. (C) The increase of K8 (panel a, d in green), K19 (panel d, f in green) and K8 pS74 (panel b, e in red arrow heads) after antibiotic-treatment was confirmed by confocal microscopy. Nuclei (DNA) are shown in blue. L = lumen, scale bar = 50 μm. (Da,b) Gene transcription analysis of K8 (Da) and K19 (Db) normalized to β-actin showed no significant changes. n.s = not significant. Numerical data are shown as average ± SD. Statistical significance for Western blot and mRNA data was determined by t-test.