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. 2023 May 25;24(11):9234. doi: 10.3390/ijms24119234

Figure 1.

Figure 1

Effect of HDM on expression of adherent and tight junctions between keratinocytes and their modification by Ceramides. (A) Primary human keratinocytes (KHN) were stimulated with HDM 100 μg/mL for 24 h and expression of adherent and tight junction proteins assessed by Western blot (WB) analysis. HSP90 was used as a loading control. (B) E-cadherin expression in the same cells stimulated with HDM in the presence or absence of Ceramide AD™, DS Phytoshingosine or DS Ceramide Y30 at 1/20, 1/50 or 1/100 dilution dissolved in 5% glycol (vehicle control). (C) WB gels were semi-quantitated by densitometric analysis using ImageJ software (version 2.1) from at least 3 separate experiments and normalized to HKG (HSP90) relative quantity (RQ) (n = 3). Numbers 1–10 denote corresponding lanes in (B). (D) Immunofluorescence staining of KHN stimulated with (bottom panels) or without (top panels) HDM 100 μg/mL in the presence of separate Ceramides whereby E-cadherin-positive cells are shown as red and DAPI-positive cells are shown as blue. * p < 0.05, *** p < 0.001 vs. respective controls and # p < 0.05 vs. HDM.