Journal: Molecules

Article Title: Substituent Effects Control the Biological Activity of Mn(II) Imidazo[1,2-a]pyridine Complexes

doi: 10.3390/molecules31061007

Figure Lengend Snippet: ( a ) Cellular uptake. Final intracellular manganese concentration expressed as ng Mn per mg protein after 24h incubation with the A549, MCF-7, PANC-1, DU-145, HT-29, and HaCat cell lines for complexes 1 , 2 , 3 in c = 1 μM. ( b ) The ROS production in PANC-1 cells after 24 h using H 2 DCF-DA for Mn compounds ( 1 , 2 , 3 ) with and without NAC (5 mM); ctrl (+): H 2 O 2 as positive control, and ctrl (−): negative control, cells without compound. ( c ) The ROS generation was monitored by H 2 DCF-DA assay in PANC-1 cells during steady incubation with Mn complexes for 30 min, 6, 12, 24, 48 and 72 h in c = 1 μM; ctrl (+): H 2 O 2 as positive control, and ctrl (−): negative control, cell without compound. ( d ) Alteration in mitochondrial membrane potential (MMP) is given as an emission ratio 570 nm/530 nm (ctrl—untreated cells; ciprofloxacin—a negative control; gentamicin—a positive control).

Article Snippet: HT-29 (colorectal adenocarcinoma, morphology: epithelial, ATCC: HTB-38), MCF7 cell line (human breast adenocarcinoma, morphology: epithelial-like, ATCC: HTB-22), A549 cell line (human lung adenocarcinoma, morphology: epithelial, ATCC: CCL-185), PANC-1 (human pancreatic/duct carcinoma, morphology: epithelial, ATCC: CRL-1469) and HaCaT (human keratinocyte, ATCC PCS-200-011) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Corning, NY, USA), supplemented with 10% fetal bovine serum (FBS) and with 1% streptomycin/penicillin.

Techniques: Concentration Assay, Incubation, Positive Control, Negative Control, Membrane

Journal: Molecules

Article Title: Substituent Effects Control the Biological Activity of Mn(II) Imidazo[1,2-a]pyridine Complexes

doi: 10.3390/molecules31061007

Figure Lengend Snippet: Cellular uptake. Final intracellular manganese concentration expressed in percentages after 24 h incubation of 1_M and 3_M (c = 0.05 mg/mL and 0.03 mg/mL, respectively) with the PANC-1 and HaCat cell lines.

Article Snippet: HT-29 (colorectal adenocarcinoma, morphology: epithelial, ATCC: HTB-38), MCF7 cell line (human breast adenocarcinoma, morphology: epithelial-like, ATCC: HTB-22), A549 cell line (human lung adenocarcinoma, morphology: epithelial, ATCC: CCL-185), PANC-1 (human pancreatic/duct carcinoma, morphology: epithelial, ATCC: CRL-1469) and HaCaT (human keratinocyte, ATCC PCS-200-011) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Corning, NY, USA), supplemented with 10% fetal bovine serum (FBS) and with 1% streptomycin/penicillin.

Techniques: Concentration Assay, Incubation

Journal: Biology Direct

Article Title: BRD4 sustains p63 transcriptional program in keratinocytes

doi: 10.1186/s13062-024-00547-1

Figure Lengend Snippet: BRD4 and p63 regulates keratinocytes proliferation and differentiation. a, b, e, f Cell cycle analysis performed in HEKn comparing SCR condition and sip63 and siBRD4 conditions comparing untreated cells (DMSO) and JQ1 treated cells (10 µM). Cells were stained with Propidium Iodide (50 µg/ml) for 1 h and then analysed by flow cytometry. In a, e representative plot of cell cycle analysis performed in HEKn is shown, while in b, f a quantification of cell cycle analysis. Graphs present means ± SD of three independent experiments. c, g EdU-incorporation assay of HEKn cells transfected with siSCR, sip63 and different BRD4 (siBRD4#1, BRD4#2) siRNAs or treated with DMSO and JQ1 (10 µM). Data are shown as mean ± SD of N = 3 experiments. (Unpaired Student’s t test). d, h Growth curve of HEKn cells transfected with siSCR, sip63 and siBRD4#1, or treated with DMSO and JQ1 (10 µM). The cell confluency has been determined using the Incucyte real-time video imaging system. Each data points indicate mean ± SEM. i – n Western blot analysis was carried out with specific antibodies against K10, and β-actin was used as loading control. ImageJ program was used to quantitate the protein levels. The blot is representative of three independent experiments. DD, days of differentiation

Article Snippet: Primary Normal Human Epidermal Keratinocytes, neonatal (HEKn) (Gibco, catalog no. C-001-5C) and human TERT-immortalized keratinocytes (Ker-CT) (ATCC, CRL4048, lot. no. 0213) were cultured in EpiLife medium with the addition of Human Keratinocyte Growth Supplements (HKGS, Life Technologies).

Techniques: Cell Cycle Assay, Staining, Flow Cytometry, Transfection, Imaging, Western Blot, Control

Journal: Biology Direct

Article Title: BRD4 sustains p63 transcriptional program in keratinocytes

doi: 10.1186/s13062-024-00547-1

Figure Lengend Snippet: Keratinocytes lacking for p63 and BRD4 have similar expression profiles. Gene expression analysis performed using nanoString technology on HEKn cells after p63 and BRD4 silencing and JQ1 treatment. a Venn diagram represents common dysregulated genes in the different conditions. b – d Pathway gene ontology of common dysregulated genes. e – g Expression levels of the shared dysregulated genes from the top five enriched categories of the GO-pathway analysis

Article Snippet: Primary Normal Human Epidermal Keratinocytes, neonatal (HEKn) (Gibco, catalog no. C-001-5C) and human TERT-immortalized keratinocytes (Ker-CT) (ATCC, CRL4048, lot. no. 0213) were cultured in EpiLife medium with the addition of Human Keratinocyte Growth Supplements (HKGS, Life Technologies).

Techniques: Expressing, Gene Expression

Journal: Scientific Reports

Article Title: Plant extract mixture shows anti-inflammatory and barrier-strengthening effects and activates aryl hydrocarbon receptor in a 2D psoriasis model

doi: 10.1038/s41598-026-50000-8

Figure Lengend Snippet: Plant extract mixture restores gene expression of skin barrier molecules filaggrin and loricrin in the 2D psoriasis model. CaCl 2 -differentiated NHEKs were left unstimulated (control) or stimulated with a psoriasis-like cytokine mixture (IL-1β, IL-17A, IL-22 and TNF-alpha, each 10 ng/mL) either in the absence (Psoriasis) or in the presence of the plant extract (Psoriasis + Extract) for 21 h. Gene expression levels of ( a ) FLG , ( b ) LOR and ( c ) IVL were determined by real-time PCR. Statistical significance was tested by a, c) one-way ANOVA with subsequent Sidak’s multiple comparison test or b) Kruskal-Wallis test with subsequent Dunn’s multiple comparisons test ( n = 12 * p < 0.05; ** p < 0.01; *** p < 0.001; ns = not significant).

Article Snippet: For the 2D psoriasis model, primary normal human epidermal keratinocytes (NHEKs, pooled from four donors, PromoCell, Heidelberg Germany), used at passage 4, were seeded in a 24-well plate in Derma Life Complete medium (CellSystems, Troisdorf, Germany).

Techniques: Plant Extract, Gene Expression, Control, Real-time Polymerase Chain Reaction, Comparison

Journal: Scientific Reports

Article Title: Plant extract mixture shows anti-inflammatory and barrier-strengthening effects and activates aryl hydrocarbon receptor in a 2D psoriasis model

doi: 10.1038/s41598-026-50000-8

Figure Lengend Snippet: Plant extract mixture downregulates inflammatory markers in the 2D psoriasis model. CaCl 2 -differentiated NHEKs were stimulated as described in Fig. . Gene expression levels of ( a ) IL1A , ( b ) IL1B, ( c ) IL1RN , ( d ) CXCL8 , ( e ) TNFA , ( f ) IL17C , ( g ) IL36G , ( h ) CSF2 , ( i ) VEGFA were measured. Statistical significance was tested by a, b, c, e, h) one-way ANOVA with subsequent Sidak’s multiple comparison test or d, f, g) Kruskal-Wallis test with subsequent Dunn’s multiple comparisons test ( n = 12, * p < 0.05; ** p < 0.01; *** p < 0.001; ns = not significant). Protein expression levels of ( j ) IL-8 and k) TNFα were measured in the supernatants of the cells by ELISA. Values for unstimulated control and plant extract-treated cells were below the detection limit of 31,3 pg/mL (IL-8) or 62,5 pg/mL (TNFα) and not detectable; therefore, no statistical analysis was performed. n.d.= non-detectable.

Article Snippet: For the 2D psoriasis model, primary normal human epidermal keratinocytes (NHEKs, pooled from four donors, PromoCell, Heidelberg Germany), used at passage 4, were seeded in a 24-well plate in Derma Life Complete medium (CellSystems, Troisdorf, Germany).

Techniques: Plant Extract, Gene Expression, Comparison, Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: Scientific Reports

Article Title: Plant extract mixture shows anti-inflammatory and barrier-strengthening effects and activates aryl hydrocarbon receptor in a 2D psoriasis model

doi: 10.1038/s41598-026-50000-8

Figure Lengend Snippet: Plant extract mixture lowers upregulated antimicrobial peptide expression in the 2D psoriasis model. CaCl 2 -differentiated NHEKs were stimulated as described in Fig. . Gene expression levels of ( a ) S100A7 , ( c ) DEFB4A and ( e ) DEFB103A were measured. Statistical significance was tested by ( a , c ) one-way ANOVA with subsequent Sidak’s multiple comparison test or ( e ) Kruskal-Wallis test with subsequent Dunn’s multiple comparisons test ( n = 12). Protein expression levels of ( b ) psoriasin and ( d ) hBD2 were measured in the supernatants of the cells by ELISA. Statistical significance was tested by b) Kruskal-Wallis test with subsequent Dunn’s multiple comparisons test ( n = 12) or d) one-way ANOVA with subsequent Sidak’s multiple comparison test ( n = 6, * p < 0.05; ** p < 0.01; *** p < 0.001; ns = not significant).

Article Snippet: For the 2D psoriasis model, primary normal human epidermal keratinocytes (NHEKs, pooled from four donors, PromoCell, Heidelberg Germany), used at passage 4, were seeded in a 24-well plate in Derma Life Complete medium (CellSystems, Troisdorf, Germany).

Techniques: Plant Extract, Expressing, Gene Expression, Comparison, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Plant extract mixture shows anti-inflammatory and barrier-strengthening effects and activates aryl hydrocarbon receptor in a 2D psoriasis model

doi: 10.1038/s41598-026-50000-8

Figure Lengend Snippet: Plant extract mixture reduces NFKBIZ and NFKBIA gene expression and IκBζ protein levels. CaCl 2 -differentiated NHEKs were stimulated as described in Fig. . Gene expression levels of ( a ) NFKBIZ and ( c ) NFKBIA were measured and statistical significance was tested by one-way ANOVA with subsequent Sidak’s multiple comparisons test ( n = 12; * p < 0.05; ** p < 0.01; *** p < 0.001; ns = not significant). ( b ) Protein expression levels of IκBζ were assessed by western blot using an IκBζ antibody. Detection of pan-actin serves as a loading control. Uncropped blots are shown in Supplementary Figure .

Article Snippet: For the 2D psoriasis model, primary normal human epidermal keratinocytes (NHEKs, pooled from four donors, PromoCell, Heidelberg Germany), used at passage 4, were seeded in a 24-well plate in Derma Life Complete medium (CellSystems, Troisdorf, Germany).

Techniques: Plant Extract, Gene Expression, Expressing, Western Blot, Control

Journal: Scientific Reports

Article Title: Plant extract mixture shows anti-inflammatory and barrier-strengthening effects and activates aryl hydrocarbon receptor in a 2D psoriasis model

doi: 10.1038/s41598-026-50000-8

Figure Lengend Snippet: Plant extract mixture activates the AhR in the 2D psoriasis model. ( a ) CaCl 2 -differentiated NHEKs were stimulated as described in Fig. . Gene expression levels of CYP1A1 were measured and statistical significance was tested by Kruskal-Wallis test with subsequent Dunn’s multiple comparisons test ( n = 12). ( b ) To determine AhR activation, NHEKs were transfected with the pGudLuc6.1 plasmid containing firefly luciferase which expression depends on AhR activation and the pGL4.74 [ hRLuc /TK] reference plasmid containing renilla luciferase. One day after transfection, cells were stimulated as described in ( a ). After cell lysis, activation of AhR was determined by measurement of relative luciferase activities. Statistical significance was tested by one-way ANOVA with subsequent Sidak’s multiple comparison test ( n = 12; ** p < 0.01; *** p < 0.001; ns = not significant).

Article Snippet: For the 2D psoriasis model, primary normal human epidermal keratinocytes (NHEKs, pooled from four donors, PromoCell, Heidelberg Germany), used at passage 4, were seeded in a 24-well plate in Derma Life Complete medium (CellSystems, Troisdorf, Germany).

Techniques: Plant Extract, Gene Expression, Activation Assay, Transfection, Plasmid Preparation, Luciferase, Expressing, Lysis, Comparison

Journal: Scientific Reports

Article Title: Plant extract mixture shows anti-inflammatory and barrier-strengthening effects and activates aryl hydrocarbon receptor in a 2D psoriasis model

doi: 10.1038/s41598-026-50000-8

Figure Lengend Snippet: Downregulation of AhR inhibits the filaggrin-inducing effects but not the anti-inflammatory effects of the plant extract in the 2D psoriasis model. NHEKs were transfected with control or AhR siRNA and stimulated as described in Fig. . Gene expression levels of ( a ) CYP1A1 , ( b ) FLG , ( c ) NFKBIZ , ( d ) TNFA , ( e ) IL36G ( f ) CXCL8 and ( g ) DEFB4A were measured. Statistical significance was determined by ( a – f ) one-way ANOVA with subsequent Sidak’s multiple comparison test or ( c ) Kruskal-Wallis with subsequent Dunn’s multiple comparison test (n = 9; * p < 0.05; ** p < 0.01; *** p 0.001; ns = not significant).

Article Snippet: For the 2D psoriasis model, primary normal human epidermal keratinocytes (NHEKs, pooled from four donors, PromoCell, Heidelberg Germany), used at passage 4, were seeded in a 24-well plate in Derma Life Complete medium (CellSystems, Troisdorf, Germany).

Techniques: Plant Extract, Transfection, Control, Gene Expression, Comparison

Journal: Scientific Reports

Article Title: Plant extract mixture shows anti-inflammatory and barrier-strengthening effects and activates aryl hydrocarbon receptor in a 2D psoriasis model

doi: 10.1038/s41598-026-50000-8

Figure Lengend Snippet: The plant extract mixture exhibits antioxidant effects in the 2D psoriasis model. CaCl 2 -differentiated NHEKs were stimulated as described in Fig. . ( a ) Gene expression levels of NQO1 were measured and statistical significance was tested by one-way ANOVA with subsequent Sidak’s multiple comparisons test ( n = 12). ( b ) Intracellular reactive oxygen species were measured by DCFDA-based assay. The mean fluorescence value of the control cells was set to 100%, and the relative intracellular ROS levels of the other samples were calculated as percentages of the control. Statistical significance was tested by Kruskal-Wallis test with subsequent Dunn’s multiple comparison test ( n = 12, * p < 0.05; ** p < 0.01; *** p < 0.001; ns = not significant). ( c ) NHEKs were transfected and stimulated as described in Fig. . Gene expression levels of NQO1 were measured and statistical significance was determined by one-way ANOVA with subsequent Sidak’s multiple comparison test ( n = 9; * p < 0.05; ns = not significant).

Article Snippet: For the 2D psoriasis model, primary normal human epidermal keratinocytes (NHEKs, pooled from four donors, PromoCell, Heidelberg Germany), used at passage 4, were seeded in a 24-well plate in Derma Life Complete medium (CellSystems, Troisdorf, Germany).

Techniques: Plant Extract, Gene Expression, Fluorescence, Control, Comparison, Transfection