Journal: International Journal of Molecular Sciences

Article Title: Anti-Inflammatory and Pro-Differentiating Properties of the Aryl Hydrocarbon Receptor Ligands NPD-0614-13 and NPD-0614-24: Potential Therapeutic Benefits in Psoriasis

doi: 10.3390/ijms22147501

Figure Lengend Snippet: NPD-0614-13 and NPD-0614-24 safety profile in different skin cell populations. ( a ) Western blot and corresponding densitometric analysis of K7 and K10 protein expression in SZ95 sebocytes treated with NPD-0614-13 (25 μM), NPD-0614-24 (25 μM) and FICZ (100 nM) for 48 h. ( b ) Percentage composition of saturated, mono- and polyunsaturated fatty acids in SZ95 sebocytes treated as above for 72 h. ( c ) Western blot and corresponding densitometric analysis of p53 and phospho-EGFR protein expression in NHKs treated as above for 48 h and 5 min, respectively. ( d ) Western blot and corresponding densitometric analysis of filaggrin and involucrin protein expression in NHKs treated as above for 48 h. ( e ) Phase-contrast analysis, western blot with corresponding densitometric analysis of tyrosinase protein expression and melanin content analysis in NHMs treated with NPD-0614-13 (25 μM), NPD-0614-24 (25 μM) and FICZ (100 nM) for 72 h and 5 days, respectively. GAPDH was used as an endogenous loading control for western blot analysis. Results are expressed as the fold change respect to untreated control cells. Data represent the mean ± SD of three independent experiments.

Article Snippet: Then sections were blocked for 15 min with 5% normal goat serum in PBS and incubated overnight at 4 °C with the following primary antibodies: anti-involucrin (1:200 in PBS) (AbCam), anti-filaggrin (1:500 in PBS) (15C10; Monosan, Sanbio, Uden, Netherlands), anti-psoriasin (S100A7) (1:300 in PBS) (Sigma), and anti-beta defensin 2 (1:1000 in PBS) (AbCam).

Techniques: Western Blot, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Anti-Inflammatory and Pro-Differentiating Properties of the Aryl Hydrocarbon Receptor Ligands NPD-0614-13 and NPD-0614-24: Potential Therapeutic Benefits in Psoriasis

doi: 10.3390/ijms22147501

Figure Lengend Snippet: NPD-0614-13 and NPD-0614-24 safety profile on human epidermal equivalents (HEEs). ( a ) Hematoxilin–eosin staining of HEEs cultured in presence or absence of NPD-0614-13, NPD-0614-24 (25 μM) and FICZ (100 nM) for 72 h. Bar: 50 μm. ( b ) Morphometric analysis of the stratum corneum (SC) and the viable epidermal layers (VE). The thickness of SC and VE was expressed as mean value ± SD (* p < 0.05 vs. untreated HEEs). ( c ) Representative immunofluorescence images after using filaggrin antibody. Nuclei were counterstained with DAPI. Bar: 20 μm. ( d ) Quantitative analysis of fluorescence intensity was expressed as mean value ± SD. Dashed black/white lines represent the junction between the basal layer and the membrane of the insert. ( e ) Quantitative real time PCR analysis of filaggrin, caspase-14, S100A7 and beta-defensin-2 in HEEs, in the presence of absence of NPD-0614-13, NPD-0614-24 (25 μM) and FICZ (100 nM) for 72 h. All mRNA values were normalized against the expression of GAPDH and were expressed relative to untreated HEEs.

Article Snippet: Then sections were blocked for 15 min with 5% normal goat serum in PBS and incubated overnight at 4 °C with the following primary antibodies: anti-involucrin (1:200 in PBS) (AbCam), anti-filaggrin (1:500 in PBS) (15C10; Monosan, Sanbio, Uden, Netherlands), anti-psoriasin (S100A7) (1:300 in PBS) (Sigma), and anti-beta defensin 2 (1:1000 in PBS) (AbCam).

Techniques: Staining, Cell Culture, Immunofluorescence, Fluorescence, Real-time Polymerase Chain Reaction, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Anti-Inflammatory and Pro-Differentiating Properties of the Aryl Hydrocarbon Receptor Ligands NPD-0614-13 and NPD-0614-24: Potential Therapeutic Benefits in Psoriasis

doi: 10.3390/ijms22147501

Figure Lengend Snippet: Effects of NPD-0614-13 and NPD-0614-24 on psoriasis-like human epidermal equivalents (PS-HEEs). ( a ) Hematoxilin–eosin staining of HEEs and PS-HEEs cultured in the presence or absence of NPD-0614-13, NPD-0614-24 (25 μM) and FICZ (100 nM) for 72 h. Bar: 50 μm. ( b ) Morphometric analysis of stratum corneum (SC) and viable epidermal layers (VE). The thickness of SC and VE was expressed as mean value ± SD (significance vs. untreated HEEs or vs. PS-HEEs are marked with * and °, respectively; ** p < 0.01 vs. untreated HEEs; p < 0.01 vs. PS-HEEs). ( c ) Representative immunofluorescence images after using antibodies against involucrin, filaggrin, S100A7, and beta-defensin-2. Nuclei were counterstained with DAPI. Bar: 20 μm. ( d ) Quantitative analysis of fluorescence intensity was expressed as mean value ± SD (* p < 0.05, ** p < 0.01 vs. untreated HEEs; ° p < 0.05, °° p < 0.01 vs. PS-HEEs). Dashed black/white lines represent the junction between the basal layer and the membrane of the insert. Quantitative real time PCR analysis of the expression of ( e ) involucrin, filaggrin, caspase-14, S100A7, beta-defensin 2, and ( f ) IL-1α, IL-1β, IL-6, and IL-8 in PS-HEEs in the presence of absence of NPD-0614-13, NPD-0614-24 (25 μM), or FICZ (100 nM) for 72 h. All mRNA values were normalized against the expression of GAPDH and were expressed relative to untreated HEEs (* p < 0.05, ** p < 0.01 vs. HEEs; p < 0.05 vs. PS-HEEs).

Article Snippet: Then sections were blocked for 15 min with 5% normal goat serum in PBS and incubated overnight at 4 °C with the following primary antibodies: anti-involucrin (1:200 in PBS) (AbCam), anti-filaggrin (1:500 in PBS) (15C10; Monosan, Sanbio, Uden, Netherlands), anti-psoriasin (S100A7) (1:300 in PBS) (Sigma), and anti-beta defensin 2 (1:1000 in PBS) (AbCam).

Techniques: Staining, Cell Culture, Immunofluorescence, Fluorescence, Real-time Polymerase Chain Reaction, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Anti-Inflammatory and Pro-Differentiating Properties of the Aryl Hydrocarbon Receptor Ligands NPD-0614-13 and NPD-0614-24: Potential Therapeutic Benefits in Psoriasis

doi: 10.3390/ijms22147501

Figure Lengend Snippet: Effects of NPD-0614-13 and NPD-0614-24 on psoriasis skin equivalents (PSEs). ( a ) Hematoxilin–eosin staining of PSEs cultured in the presence or absence of NPD-0614-13, NPD-0614-24 (25 μM) and FICZ (100 nM) for 4 days. Bar: 50 μm. ( b ) Morphometric analysis of stratum corneum (SC) and viable epidermal layers (VE). The thickness of SC and VE was expressed as mean value ± SD (* p < 0.05, ** p < 0.01 vs. PSEs). ( c ) Representative immunofluorescence images after using antibodies against filaggrin and beta-defensin-2. Nuclei were counterstained with DAPI. Bar: 50 μm. ( d ) Quantitative analysis of fluorescence intensity was expressed as mean value ± SD (** p < 0.01 vs. PSEs). ( e ) IL-6 and IL-8 quantitation by ELISA in PSEs in the presence of absence of NPD-0614-13, NPD-0614-24 (25 μM) or FICZ (100 nM) for 72 h. The data are expressed as mean ± SD (** p < 0.01 vs. PSEs).

Article Snippet: Then sections were blocked for 15 min with 5% normal goat serum in PBS and incubated overnight at 4 °C with the following primary antibodies: anti-involucrin (1:200 in PBS) (AbCam), anti-filaggrin (1:500 in PBS) (15C10; Monosan, Sanbio, Uden, Netherlands), anti-psoriasin (S100A7) (1:300 in PBS) (Sigma), and anti-beta defensin 2 (1:1000 in PBS) (AbCam).

Techniques: Staining, Cell Culture, Immunofluorescence, Fluorescence, Quantitation Assay, Enzyme-linked Immunosorbent Assay