normal human epidermal keratinocytes nhek  (PromoCell)

Journal: Communications Biology

Article Title: The MYB-related transcription factor MYPOP acts as a selective regulator of cancer cell growth

doi: 10.1038/s42003-026-10272-2

Figure Lengend Snippet: a MYPOP expression in normal (N) and tumor (T) cells detected by Western blotting showing endogenous protein levels of MYPOP in normal human epidermal keratinocytes (NHEK) and cervical cancer cells (HeLa). In addition, the expression of GFP and MYPOP in HeLa cells after transfection with pcDNA3.1-MYPOP (MYPOP), pEGFP-C3-MYPOP (GFP-MYPOP) or corresponding controls (pcDNA3.1, Control and pEGFP-C3, GFP-Control) is shown. MYPOP and GFP were stained using anti-MYPOP pAb and anti-GFP mAb. GAPDH staining was used as a loading control. b – i GFP-Control or GFP-MYPOP expressing HeLa cells. b , c Lower ( b ) and higher ( c ) magnification for representative fluorescence microscopy images of GFP-Control or GFP-MYPOP (green) expressing cells 24 h post transfection (p.t.). Cell nuclei were stained using Hoechst 33342 (blue). d , e Representative fluorescence microscopy images treated as in ( c ) showing cytoplasmic, nucleocytoplasmic ( d ) and nuclear ( e ) localization as well as co-localization of GFP-MYPOP (green) and DNA (blue). f Co-localization analysis between GFP or GFP-MYPOP and DNA (Hoechst) using Pearson correlation coefficient (PCC). At least 10 GFP-positive cells were analyzed for each treatment and biological replicate ( n = 3). Values are shown as mean + SD. Statistical significance was determined with p = 0.0001 comparing GFP-MYPOP and GFP-Control. g Representative fluorescence microscopy images of GFP-MYPOP (green) expressing cells showing shrunken or fragmented nuclei at 24 h p.t. Cell nuclei (blue) as above. h , i Quantification of fragmented and shrunken nuclei at 24 h p.t ( h ) and 48 h p.t ( i ). At least 100 GFP-positive cells were analyzed for each treatment, time point and biological replicate, respectively. Values ( n = 3) are shown as mean + SD. Statistical significance was determined with p (24 h p.t.) = 0.0036 and p (48 h p.t.) = 0.0428 comparing GFP-MYPOP and GFP-Control.

Article Snippet: Normal Human Epidermal Keratinocytes (NHEK) were purchased from PromoCell, Germany and were cultivated according to the manufacturer’s instructions.

Techniques: Expressing, Western Blot, Transfection, Control, Staining, Fluorescence, Microscopy

Journal: Communications Biology

Article Title: The MYB-related transcription factor MYPOP acts as a selective regulator of cancer cell growth

doi: 10.1038/s42003-026-10272-2

Figure Lengend Snippet: a Endogenous protein levels of MYPOP in untreated cancer cell lines. Normal skin cells (NHEK) and normal lung cells (181576 N, 181652 N) served as controls. GAPDH or β-actin staining was used as loading control. b Cancer cells were transfected with either a MYPOP expression plasmid or a control plasmid, selected for 6–12 days with G418, fixed, and stained with crystal violet. The cell-covered area was quantified (relative area). Values are presented as mean + SD. The mean for control-transfected cells was set to 100% (dotted line). Statistical significance was determined by comparing control and MYPOP expressing cells with p = 0.0022 for Huh7 ( n = 3), p = 0.0028 for HEK293 ( n = 5), p = 0.0064 for MCF7 ( n = 4), p = 0.0294 for HCT116 ( n = 3), p = 0.0106 for HeLa ( n = 3), p = 0.0003 for A549 ( n = 3), and p = 0.0026 for 2106 T ( n = 4).

Article Snippet: Normal Human Epidermal Keratinocytes (NHEK) were purchased from PromoCell, Germany and were cultivated according to the manufacturer’s instructions.

Techniques: Staining, Control, Transfection, Expressing, Plasmid Preparation

Journal: Communications Biology

Article Title: The MYB-related transcription factor MYPOP acts as a selective regulator of cancer cell growth

doi: 10.1038/s42003-026-10272-2

Figure Lengend Snippet: a Protein expression of untreated, control mRNA and MYPOP mRNA-transfected HeLa cells and NHEK cells, as indicated at different time points post transfection (6 h–54 h) was analyzed by western blotting using anti-MYPOP and anti-GAPDH antibodies. Under conditions of low protein loading and short exposure times, endogenous MYPOP is not detected in NHEK, as these parameters are optimized to prevent oversaturation of overexpressed MYPOP at 6 h p.t. Detection of endogenous MYPOP increases at later time points, reflecting cell growth and higher total protein content. Lower panel: optical microscope overview images of untreated, control mRNA and MYPOP mRNA-transfected HeLa cells (left panel) and NHEK cells (right panel) at 6 h and 54 h after mRNA transfection. b Growth curves (object counts per image, measurement every 2 h) of untreated, control mRNA and MYPOP mRNA-transfected HeLa, NHEK, CaSki, HaCaT, HCT116, and 2106 T cells at the indicated time points. Statistical significance ( n = 4 for HeLa, n = 3 for all others) was determined between Control and MYPOP cell counts at 70 h or 72 h p.t. as indicated with p = 0.0203 for HeLa, p = 0.0441 for CaSki, p = 0.4720 for HCT116, p = 0.2424 for NHEK, p < 0.0001 for HaCaT, and p = 0.0152 for 2106 T. c Left: MYPOP and GAPDH protein expression in Hela wild-type (WT) and MYPOP knockout (KO) cells. Statistical significance ( n = 3) between WT and KO cells was determined with p = 0.0014 for MYPOP band intensities. Center: optical microscope overview images of HeLa WT and KO cells. Right: growth curve of HeLa WT and KO cells. Statistical significance ( n = 4) was determined with p = 0.6099 at 72 h t.p.

Article Snippet: Normal Human Epidermal Keratinocytes (NHEK) were purchased from PromoCell, Germany and were cultivated according to the manufacturer’s instructions.

Techniques: Expressing, Control, Transfection, Western Blot, Microscopy, Knock-Out

Journal: Communications Biology

Article Title: The MYB-related transcription factor MYPOP acts as a selective regulator of cancer cell growth

doi: 10.1038/s42003-026-10272-2

Figure Lengend Snippet: Volcano plots depicting gene expression changes of control and MYPOP mRNA-transfected HeLa and NHEK cells at 6 h and 24 h p.t. as indicated. The labels display the overlapping DEGs, which were selected from the initial RNA-Seq experiment shown in Fig. , comprising the 30 top DEGs (15 up, 15 downregulated) and the 27 ‘cell cycle’ genes. Significantly downregulated candidates with adjusted p ≤ 0.05 are shown in blue, and upregulated candidates with adjusted p ≤ 0.05 are shown in red.

Article Snippet: Normal Human Epidermal Keratinocytes (NHEK) were purchased from PromoCell, Germany and were cultivated according to the manufacturer’s instructions.

Techniques: Gene Expression, Control, Transfection, RNA Sequencing

Journal: Applied and Environmental Microbiology

Article Title: Skin Staphylococcus species differentially modulate keratinocyte cytokine secretion in response to UVB

doi: 10.1128/aem.01549-25

Figure Lengend Snippet: Dose-dependent effects on cytokine secretion. NHEKs (p3–5) cultured to 80% confluency were irradiated at 17–33 mJ/cm². Supernatants were collected 24 h post-irradiation and assessed via cytokine array ( A ) or via ELISA for IL-6 ( B ), IL-8 ( C ), CCL20 ( D ), and TNF-α ( E ). ELISA data are presented as mean ± SD. **** P < 0.0001, *** P < 0.001, ** P < 0.01, and * P < 0.05, as determined via one-way ANOVA followed by Tukey test.

Article Snippet: Pooled ( n = 3), normal human epidermal keratinocytes (NHEKs from juvenile foreskin, Promocell C-12005; passages 1–5) were routinely cultured in keratinocyte growth medium (KGM-2, Promocell).

Techniques: Cell Culture, Irradiation, Enzyme-linked Immunosorbent Assay

Journal: Applied and Environmental Microbiology

Article Title: Skin Staphylococcus species differentially modulate keratinocyte cytokine secretion in response to UVB

doi: 10.1128/aem.01549-25

Figure Lengend Snippet: Exposure to a skin PMC elevates cytokine secretion regardless of UVB exposure. NHEKs (p2–5) were treated with PBS (PBS, no UVB) or S. epidermidis, S. hominis, M. luteus, C. tuberculostearicum, and C. acnes for 30–90 min and subsequently exposed to UVB (33 mJ/cm²). Supernatants were collected 24 h post-inoculation with each organism, and the effects of the skin microbial PMC ± UVB on IL-6 ( A ), IL-8 ( B ), TNF-α ( C ), and CCL20 ( D ) secretion were assessed using ELISA. ** P < 0.01 and * P < 0.05, as determined via one-way ANOVA followed by Tukey test.

Article Snippet: Pooled ( n = 3), normal human epidermal keratinocytes (NHEKs from juvenile foreskin, Promocell C-12005; passages 1–5) were routinely cultured in keratinocyte growth medium (KGM-2, Promocell).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Applied and Environmental Microbiology

Article Title: Skin Staphylococcus species differentially modulate keratinocyte cytokine secretion in response to UVB

doi: 10.1128/aem.01549-25

Figure Lengend Snippet: UVB impacts the relative abundance of skin commensals in a polymicrobial skin community. NHEKs (p2–5) were incubated with a polymicrobial community for 90 min, media replaced, and cultures exposed to 33 mJ/cm2 UVB or left unirradiated (no UVB). Polymicrobial community composition was assessed 24 h post-treatment via qPCR, where relative proportions of each organism were calculated via species-specific standard curves and presented as collated data ( A , results for C. tuberculostearicum and M. luteus are not visible in the graph, as abundance was <1% of the total community) and UVB induced changes on individual species within the community of S. epidermidis ( B ), S. hominis ( C ), C. acnes ( D ), M. luteus ( E ), and C. tuberculostearicum ( F ). Data are presented as mean ± SD. ** P < 0.01 and * P < 0.05, as determined via unpaired t -test.

Article Snippet: Pooled ( n = 3), normal human epidermal keratinocytes (NHEKs from juvenile foreskin, Promocell C-12005; passages 1–5) were routinely cultured in keratinocyte growth medium (KGM-2, Promocell).

Techniques: Incubation

Journal: Applied and Environmental Microbiology

Article Title: Skin Staphylococcus species differentially modulate keratinocyte cytokine secretion in response to UVB

doi: 10.1128/aem.01549-25

Figure Lengend Snippet: UVB exacerbates S. epidermidis -induced cytokine secretion, while S. hominis amplified secretion of IL-8 and CCL20 regardless of UVB exposure. NHEKs (p2–5) were incubated with media alone, S. hominis, or S. epidermidis for 30–60 min, media replaced with PBS, and exposed to UVB (0–33 mJ/cm²). Adherent bacteria were then co-cultured with NHEKs for 24 h, following which supernatants were collected to assess the effects on IL-6 ( A and E ), CCL20 ( B and F ), IL-8 ( C and G ), and TNF-α ( D and H ). Significance was assessed via one-way ANOVA followed by Tukey test, where ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: Pooled ( n = 3), normal human epidermal keratinocytes (NHEKs from juvenile foreskin, Promocell C-12005; passages 1–5) were routinely cultured in keratinocyte growth medium (KGM-2, Promocell).

Techniques: Amplification, Incubation, Bacteria, Cell Culture

Journal: Applied and Environmental Microbiology

Article Title: Skin Staphylococcus species differentially modulate keratinocyte cytokine secretion in response to UVB

doi: 10.1128/aem.01549-25

Figure Lengend Snippet: S. hominis diminishes S. epidermidis- enhanced cytokine responses to UVR. NHEKs (p2–5) were incubated with media, S. hominis , S. epidermidis, or both for 30–60 min, media replaced with PBS, and subsequently exposed to UVB (0–20 mJ/cm²). Twenty-hours post-microbial inoculation, supernatants were collected and effects on cytokine secretion assessed by ELISA ( A–D ). **** P < 0.0001, *** P < 0.001, ** P < 0.01, and * P < 0.05, as determined via one-way ANOVA followed by Tukey test.

Article Snippet: Pooled ( n = 3), normal human epidermal keratinocytes (NHEKs from juvenile foreskin, Promocell C-12005; passages 1–5) were routinely cultured in keratinocyte growth medium (KGM-2, Promocell).

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: Biomolecules

Article Title: Mandelamide Isolated from Prunus persica Flowers Attenuates TNF-α–Driven Oxidative and Inflammatory Responses in Human Skin Cells

doi: 10.3390/biom16050672

Figure Lengend Snippet: Effects of mandelamide ( 4 ) on pro-inflammatory mediator secretion in TNF-α–stimulated human epidermal keratinocytes (HEKs). Cells were pretreated with mandelamide at the indicated concentrations (12.5, 25, 50, and 100 μM) for 1 h, followed by stimulation with TNF-α for 12 or 24 h. The secretion levels of IL-6, IL-8, and IL-1β ( A ), as well as COX-2, PGE 2 , and nitric oxide (NO) ( B ), were quantified using ELISA, while NO production was additionally measured using the Griess assay. All experiments were performed according to the manufacturers’ protocols. Data are presented as mean ± SEM ( n = 3). Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test; # p < 0.05, ## p < 0.01, and ### p < 0.001 vs. control; * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. TNF-α–treated group. Dexamethasone (Dexa) was used as a positive control in the IL-6 and IL-8 ELISAs.

Article Snippet: Human dermal fibroblasts (HDFs; CAT No. C-12302, adult donor) and human epidermal keratinocytes (HEKs; CAT No. C-12006, pooled adult donor) were purchased from PromoCell GmbH (Heidelberg, Germany).

Techniques: Enzyme-linked Immunosorbent Assay, Griess Assay, Control, Positive Control

Journal: Biomolecules

Article Title: Mandelamide Isolated from Prunus persica Flowers Attenuates TNF-α–Driven Oxidative and Inflammatory Responses in Human Skin Cells

doi: 10.3390/biom16050672

Figure Lengend Snippet: Effects of mandelamide ( 4 ) on the mRNA expression of MMPs and collagen-related genes in TNF-α/IFN-γ–stimulated human epidermal keratinocytes (HEKs). Cells were pretreated with mandelamide at the indicated concentrations (12.5, 25, 50, and 100 μM) for 1 h, followed by stimulation with TNF-α/IFN-γ for 24 h. The mRNA expression levels of MMP-1, MMP-2, MMP-9 ( A ), as well as COL1A1, COL1A2, COL3A1, and COL4A1 ( B ) were quantified by quantitative real-time PCR (qRT-PCR). All experiments were performed according to the manufacturers’ protocols. Data are presented as mean ± SEM ( n = 3). Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test; ## p < 0.05 and ### p < 0.001 vs. control; *, **, and *** p < 0.05, p < 0.01, and p < 0.001 vs. TNF-α/IFN-γ–treated group.

Article Snippet: Human dermal fibroblasts (HDFs; CAT No. C-12302, adult donor) and human epidermal keratinocytes (HEKs; CAT No. C-12006, pooled adult donor) were purchased from PromoCell GmbH (Heidelberg, Germany).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Control

Journal: Biomolecules

Article Title: Mandelamide Isolated from Prunus persica Flowers Attenuates TNF-α–Driven Oxidative and Inflammatory Responses in Human Skin Cells

doi: 10.3390/biom16050672

Figure Lengend Snippet: Effects of mandelamide ( 4 ) on hyaluronan synthase (HAS) gene expression in TNF-α/IFN-γ–stimulated human epidermal keratinocytes (HEKs). Cells were pretreated with mandelamide at the indicated concentrations (12.5, 25, 50, and 100 μM) for 1 h, followed by stimulation with TNF-α/IFN-γ for 24 h. The mRNA expression levels of HAS-1, HAS-2, and HAS-3 were quantified by quantitative real-time PCR (qRT-PCR). All experiments were performed according to the manufacturers’ protocols. Data are presented as mean ± SEM ( n = 3). Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test; ## p < 0.05 and ### p < 0.001 vs. control; *, **, and *** p < 0.05, p < 0.01, and p < 0.001 vs. TNF-α/IFN-γ–treated group.

Article Snippet: Human dermal fibroblasts (HDFs; CAT No. C-12302, adult donor) and human epidermal keratinocytes (HEKs; CAT No. C-12006, pooled adult donor) were purchased from PromoCell GmbH (Heidelberg, Germany).

Techniques: Gene Expression, Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, 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 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

Journal: bioRxiv

Article Title: Comparative analysis of varicella-zoster virus and herpes simplex virus 1 interaction with epidermal terminal differentiation in primary human keratinocytes models of differentiation

doi: 10.64898/2026.04.08.717198

Figure Lengend Snippet: A. Schematic of epidermal differentiation and the differentiation stages at which viral infections were implemented; undifferentiated (Undiff.), early differentiated (Early diff.) and late differentiated (Late diff.). B. Schematic of plan of infection with VZV at MOI 0.1 of Undiff., Early diff. and Late diff. NHEKs. “Undiff.”, “Early diff.” and “Late diff.” labelling refers to the state of NHEKs differentiation at the time of infection (day 0). The “Undiff.” condition includes the NHEKs undergoing differentiation by calcium switch after infection (Differentiating_sync.) C. Analysis by qPCR of cell-associated VZV genome copy number normalised to number of cells, upon infection with VZV of Undiff., Early diff. and Late diff. NHEKs, and evaluated at 24h and 120h p.i. The data are generated from quantification of the VZV ORF31 gene (gB) normalised to RPP30 gene (host internal control) and are reported as the mean of three technical replicates. D. Analysis by qPCR of cell-associated VZV genome copy number normalised to number of cells upon infection with VZV of Late diff. NHEKs, and evaluated at 5 days (120h) and 7 days p.i. The data are generated from quantification of the VZV ORF31 gene (gB) normalised to RPP30 gene (host internal control) and are reported as the mean of three technical replicates. E. Analysis by qRT-PCR of ORFC2 (encoding the IE62 protein) and ORFC8 (encoding the gE protein) in VZV infection of Undiff., Early diff. and Late diff. NHEKs, and reported at 24h and 120h p.i. The data are reported as mean of 2 -dCT of three technical replicates, where the normalisation was performed against GAPDH . F. Analysis by qRT-PCR of ORFC2 and ORFC8 in VZV infection of Late diff. NHEKs, and evaluated at 120h and 7 days p.i. The data are reported as mean of 2 -dCT of three technical replicates, where the normalisation was performed against GAPDH . G. Western Blotting analysis of gE and IE62 expression at 120h p.i. following infection of Undiff., Early diff. and Late diff. NHEKs with VZV. GAPDH was used as loading control . H. Fast red staining of gE at 120h p.i. following infection of Undiff., Early diff. and Late diff. NHEKs with VZV. Graph shows the quantification of number of VZV plaques in at least two fields of view per condition and is reported as mean ± SD. I. IE62 and gE (green) immunofluorescence staining of Undiff. and Early diff. NHEKs infected with VZV and analysed at 120h p.i. DAPI (blue) was used for staining of nuclei. Graphs show quantification of integrated density of IE62 and gE signals in at least two fields of view per condition ± SD. Data in C., E. and G. are representative of n=3 independent experiments (for the Undiff. and Early Diff. conditions), data in I. are representative of n=2 independent experiments. Statistical significance was evaluated in H. by one-way ANOVA with Dunnett’s multiple comparisons test, in I. by two-tailed t test. Statistical significance is indicated as *P< 0.05, **P< 0.01, ***P< 0.001. Scale bar, 50 μm. Undiff., undifferentiated; Early diff., early differentiated; Late diff., late differentiated; p.i., post infection; NHEKs, normal human epidermal keratinocytes.

Article Snippet: Primary normal human epidermal keratinocytes (NHEKs) isolated from juvenile foreskin and pooled from different donors were purchased from Promocell or Thermo Fisher Scientific and grown are previously described [ , ].

Techniques: Infection, Generated, Control, Quantitative RT-PCR, Western Blot, Expressing, Staining, Immunofluorescence, Two Tailed Test

Journal: bioRxiv

Article Title: Comparative analysis of varicella-zoster virus and herpes simplex virus 1 interaction with epidermal terminal differentiation in primary human keratinocytes models of differentiation

doi: 10.64898/2026.04.08.717198

Figure Lengend Snippet: A. Schematic of plan of infection with HSV-1 at MOI 0.005 of undifferentiated (Undiff.), early differentiated (Early diff.) and late differentiated (Late diff.) NHEKs. “Undiff.”, “Early diff.” and “Late diff.” labelling refers to the state of NHEKs differentiation at the time of infection (day 0). The “Undiff.” condition includes the NHEKs undergoing spontaneous asynchronous differentiation after infection (Differentiating_async.) B. Analysis by qPCR of cell-associated HSV-1 genome copy number normalised to number of cells, upon infection with HSV-1 of Undiff., Early diff. and Late diff. NHEKs, and evaluated at 24h, 48h and 72h p.i. The data are generated from quantification of the HSV-1 UL27 gene (gB) normalised to RPP30 gene (host internal control) and are reported as mean of n=3 independent experiments ± SEM. C. Analysis by qPCR of cell-associated HSV-1 genome copy number normalised to number of cells, upon infection with HSV-1 of Late diff. NHEKs, and reported at 72h and 120h p.i. The data are generated from quantification of the HSV-1 UL27 gene (gB) normalised to RPP30 gene (host internal control) and are reported as the mean of three technical replicates. D. ICP0 (pink) immunofluorescence staining of Undiff., Early diff. and Late diff. NHEKs infected with HSV-1 and analysed at 4h, 24h and 72h p.i. The reported images include GFP (green) signal deriving from the GFP protein tagged to the late HSV-1 tegument protein UL46 and DAPI (grey) staining of nuclei. E. Analysis by qRT-PCR analysis of RL2 gene (encoding the ICP0 protein) and UL48 gene (encoding the VP16 protein) expression in HSV-1 infection of Undiff., Early diff. and Late diff. NHEKs, and reported at 24h, 48h and 72h p.i. The data are reported as mean of 2 -dCT of n=3 independent experiments ± SEM, where the normalisation was performed against GAPDH . F. Western Blotting analysis of ICP0 and VP16 expression at 24h, 48h and 72h p.i. following infection of Undiff., Early diff. and Late diff. NHEKs with HSV-1. GAPDH was used as loading control. Data are representative of n=4 independent experiments (Undiff. condition), n=3 independent experiments (Early diff. condition) and n=2 independent experiments (Late diff. condition). G. Schematic of the model of HSV-1 infection implemented in undifferentiated NHEKs, which either remained undifferentiated throughout the infection and until the last time point (72h) (Undifferentiated) or started undergoing spontaneous differentiation during the last time points of infection (Differentiating_async). H. Analysis by qRT-PCR of KRT5 and KRT10 mRNAs in uninfected NHEKs at 72h confirming the undifferentiated and differentiating status of the keratinocytes. The data are reported as fold change (FC) (2 -ddCT ) to the undifferentiated condition and are the mean of three technical replicates, where the normalisation was performed against GAPDH . I. Keratinocytes where infected when undifferentiated and then either maintained their undifferentiated status throughout the infection (Undifferentiated) or were allowed to spontaneously differentiate by 72h p.i. (Differentiating). Cell-associated HSV-1 genome copy number was analysed at 72h p.i. in both conditions. The data are generated from quantification of the HSV-1 UL27 gene (gB) normalised to RPP30 gene (host internal control), are reported as the mean of three technical replicates and are representative of n=3 independent experiments. J. Anlysis by qRT-PCR of RL2 and UL48 mRNAs in HSV-1 infected Undifferentiated and Differentiating NHEKs at 72h p.i. The data are reported as mean of 2 -dCT of three technical replicates, where the normalisation was performed against GAPDH . Data are representative of n=3 independent experiments. Statistical significance was evaluated in B. and E. by 2way ANOVA with Tukey’s multiple comparisons test and indicated as *P< 0.05, **P< 0.01, ***P< 0.001, ****P< 0.0001. In D. , dotted lines mark nuclei at 4h p.i. Insets contain zoomed-in areas of the ICP0 signal inside the nuclei at 4h p.i. Scale bar, 10 μm. Undiff., undifferentiated; Early diff., early differentiated; Late diff., late differentiated; p.i., post infection; NHEKs, normal human epidermal keratinocytes.

Article Snippet: Primary normal human epidermal keratinocytes (NHEKs) isolated from juvenile foreskin and pooled from different donors were purchased from Promocell or Thermo Fisher Scientific and grown are previously described [ , ].

Techniques: Infection, Generated, Control, Immunofluorescence, Staining, Quantitative RT-PCR, Expressing, Western Blot

Journal: bioRxiv

Article Title: Comparative analysis of varicella-zoster virus and herpes simplex virus 1 interaction with epidermal terminal differentiation in primary human keratinocytes models of differentiation

doi: 10.64898/2026.04.08.717198

Figure Lengend Snippet: A. Analysis of qRT-PCR expression of KRT10 (encoding the K10 protein) in VZV infection (at the MOI of 0.1) of Undiff., Early diff. and Late diff. NHEKs, and reported at 24h, 120h p.i. and at 7 days p.i. (in the Late diff.). The data are reported as fold change (FC) (2 -ddCT ) to each uninfected control from the mean of three technical replicates, where the normalisation was performed against GAPDH . Similar trends were observed in other 2 independent experiments for the Undiff. and Early Diff. conditions using different MOIs. B. Western Blotting analysis of K10 expression at 120h p.i. following infection of Undiff., Early diff. and Late diff. NHEKs with VZV at the MOI of 0.1. GAPDH was used as loading control. Numbers at the bottom of the blots indicate K10 densitometry expressed as fold change to each uninfected control, following normalisation against GAPDH . Data are representative of n=3 independent experiments for the Undiff. and Early Diff. conditions. C. K10 (red) and gE (green) immunofluorescence staining of Undiff., Early diff. and Late diff. NHEKs infected with VZV at the MOI of 0.1 and analysed at 120h p.i. DAPI (blue) was used for staining of nuclei. Data are representative of n=2 independent experiments for the Undiff. and Early Diff. conditions. D. Analysis by qRT-PCR of KRT10 mRNA expression in HSV-1 infection (at the MOI of 0.005) of Undiff., Early diff. and Late diff. NHEKs, and reported at 24h, 48h and 72h p.i. The data are reported as fold change (FC) (2 -ddCT ) to each uninfected control from the mean of n=3 independent experiments ± SEM, where the normalisation was performed against GAPDH . E. Western Blotting analysis of K10 expression at 24h, 48h and 72h p.i. following infection of Undiff., Early diff. and Late diff. NHEKs with HSV-1 at the MOI of 0.005. GAPDH was used as loading control. Data are representative of n=4 independent experiments for the Undiff. condition, n=3 independent experiments for the Early diff. condition and n=2 independent experiments for the Late diff. condition. In the Undiff. and Early diff. conditions 16μg of proteins were run. In the Late diff. condition 5μg of proteins were run to compensate for the high concentration of K10 in this condition. F. K10 (yellow) and VP16 (pink) immunofluorescence staining, of Undiff., Early diff. and Late diff. NHEKs infected with HSV-1 at the MOI of 0.005, and analysed at 72h p.i. DAPI (blue) was used for staining of nuclei. G. Analysis by qRT-PCR at 72h p.i. of expression of KRT10 mRNA in HSV-1 infection (at the MOI of 0.005) of Undiff. NHEKs treated either with foscarnet or with vehicle alone at the time of infection. The data are reported as fold change (FC) (2 -ddCT ) to each uninfected control, from the mean of three technical replicates, where the normalisation was performed against GAPDH . The data are representative of n=2 independent experiments. H. Keratin 10 (green) and gE (red) immunofluorescence staining of ex-vivo human skin tissues infected with VZV (10 4 PFU) and analysed 11 days p.i. DAPI (blue) was used for staining of nuclei. I. Keratin 10 (red) immunofluorescence staining of ex-vivo human skin tissues infected with HSV-1 (10 4 PFU) and analysed 11 days p.i. GFP signal (green) marks the expression of the GFP-tagged late HSV-1 protein UL46. DAPI (blue) was used for staining of nuclei. In H. and I. dotted lines indicate the epidermal-dermal junction. Scale bars, 50 μm. Undiff., undifferentiated; Early diff., early differentiated; Late diff., late differentiated; p.i., post infection; UNINF, uninfected.

Article Snippet: Primary normal human epidermal keratinocytes (NHEKs) isolated from juvenile foreskin and pooled from different donors were purchased from Promocell or Thermo Fisher Scientific and grown are previously described [ , ].

Techniques: Quantitative RT-PCR, Expressing, Infection, Control, Western Blot, Immunofluorescence, Staining, Concentration Assay, Ex Vivo

Journal: bioRxiv

Article Title: Rhomboid protease RHBDL2 is a calcium-activated suppressor of EGFR signalling in keratinocytes

doi: 10.64898/2026.03.19.712941

Figure Lengend Snippet: A. Western blot showing knockdown of endogenous RHBDL2 protein in HaCaT cells by multiple shRNAs (01, 00, 02) compared with wild-type (WT) and vector control (V) cells. B. Statistics of the duplicate proteomics experiments. Numbers of identified and quantified proteins ranked by their topology are shown for each of the SILAC experiments, their overlap and union. Type I membrane proteins (with a signal peptide and a single transmembrane helix), which are potential rhomboid substrates, represent about 25% of the secretome in each case and are shown in pale green. Blue, type II membrane proteins; turquoise, polytopic transmembrane proteins; grey, secreted proteins; black, intracellular proteins. C. Changes in membrane protein abundance in HaCaT keratinocyte secretome induced by RHBDL2 expression. In two independent reverse experiments, WT and R2kd HaCaT cells were isotopically labelled by heavy or light lysine and arginine, the media from both populations were pooled and the lectin-enriched glycoproteins were identified and quantified by MS analysis. The abundance ratios of all transmembrane proteins identified in both experiments (i.e. the overlap of the two datasets) were plotted against each other. Two-fold enrichment was set as a significance threshold (dotted line). Membrane proteins occurring in the grey quadrant showed consistent enrichment in both experiments and represent strong candidates for RHBDL2 substrates. D. HaCaT cells were stably transfected with constructs encoding fluorescent fusions of EGFR (GFP) and RHBDL2 (mCherry) and analysed by confocal microscopy. Scale bar = 10 µm. E. Media from WT, vector control (V) and RHBDL2 knockdown (R2kd) HaCaT and MDA-MB-468 cells was concentrated and probed with an antibody raised against the EGFR ectodomain to detect RHBDL2 dependent shedding at endogenous levels of expression. F. Conditioned media from HaCaT cells were divided equally and one half was subjected to high-speed ultracentrifugation (UC) to remove membranes including exosomes. The supernatant after ultracentrifugation and the untreated medium were immunoblotted using separate primary antibodies raised against the extracellular N-terminal (NT) or the intracellular C-terminal part (CT) of EGFR. Ultracentrifugation selectively depletes the full-length form of EGFR, which is reactive against the C-terminal antibody. G . Contribution of metalloproteases to EGFR shedding. N-terminally Strep-tagged EGFR was expressed in HEK293ET cells alongside HA-tagged RHBDL2 or a catalytically inactive mutant (S187A) and cultivated for 24 hrs in the presence or absence of 10 µM BB94. H. The candidate cleavage sites were identified by mass spectrometry (MS) of the purified ectodomain ( Fig. S1 ). Candidate P1 residues were mutated to proline to produce uncleavable mutants and tested by co-overexpression with the WT or inactive mutant enzyme (S187A) in HEK293ET cells. I. An RHBDL2 dependent C-terminal EGFR fragment (open arrow) can be produced by overexpression of the wild-type enzyme in HaCaT cells, but not the S187T inactive mutant (left). RHBDL2 overexpressing cells were incubated overnight with 10 nM PR-171 (PR), 10 µM lactacystin (LC), 10 µM chloroquine (CQ) or 100 nM bafilomycin A1 (BA1) to determine the fate of the fragment (middle). The fragment can also be observed by overnight treatment with bafilomycin A1 at endogenous levels of RHBDL2 expression (right).

Article Snippet: To examine the biological relevance of this mechanism, we measured RHBDL2 activity and EGFR shedding in primary Normal Human Adult Epidermal Keratinocytes (NHEK-Ad) and hTERT-immortalized keratinocytes Ker-CT (ATCC CRL-4048), more stably accessible alternatives to primary keratinocytes.

Techniques: Western Blot, Knockdown, Plasmid Preparation, Control, Multiplex sample analysis, Membrane, Quantitative Proteomics, Expressing, Stable Transfection, Transfection, Construct, Confocal Microscopy, Mutagenesis, Mass Spectrometry, Purification, Over Expression, Produced, Incubation

Journal: bioRxiv

Article Title: Rhomboid protease RHBDL2 is a calcium-activated suppressor of EGFR signalling in keratinocytes

doi: 10.64898/2026.03.19.712941

Figure Lengend Snippet: A-C . HaCaT cell lines were seeded equally and grown to confluent monolayers over 48 h. Cells were lysed on ice in the presence of protease and phosphatase inhibitors and samples were diluted to equal concentrations of total protein. Lysates were then probed for various components of EGFR signalling pathways by western blotting. D. HaCaT cell migration was analysed by time-lapse phase-contrast microscopy over the indicated timeframe. Left panel shows the time sequence of the cell colony outline during spreading of wild-type, R2kd and vector transfected cells. Colony outlines were superimposed from first image to last image. For clarity, outlines corresponding to initial 340 min are shown in 20 min increments. Right panel shows quantification of average distance migrated after 16 h. Asterisks correspond to P values (ns=P > 0.05, *=P ≤ 0.05,**= P ≤ 0.01, ***=P ≤ 0.001 and ****= P ≤ 0.0001). E. Gap-closure assay. WT, R2kd and vector HaCaT cells were grown to confluence around removable silicone inserts. After insert removal, gap closure was observed by time-lapse microscopy over 16 h. The average distance migrated is quantified in the box and whisker plot. F. Quantification of migration of WT-HaCaT, R2kd cells and rescue cells in which shRNA refractory wild type RHBDL2 (R2kd+WT) or its inactive S187T mutant (R2kd+mut) were re-introduced into R2kd cells. RHBDL2 overexpression was induced by 5 µg/mL cumate where indicated. G. Depletion of RHBDL2 potentiates invasion of HaCaT cells into a 3D collagen matrix. Spheroids of HaCaT keratinocytes, wild-type (WT), Vector control (V), RHBDL2 knockdown (R2kd) and rescue cells (R2kd+WT, R2kd+mut) were embedded in collagen and their invasion was measured after 72 h by comparing the total area of invaded cells relative to the area of the cell spheroid at 0 h. Invasion is quantified from 5-8 spheroids in each of 3 replicate experiments (total ≥20). Statistical analyses by Tukey’s multiple comparisons test were performed using Prism software (GraphPad Software Inc.). H. HaCaT cell proliferation rate after 48 h growth in 3D collagen was assayed using the Alamar Blue assay. The level of fluorescence is proportional to the metabolic activity and hence can be used to estimate the number of cells relative to each line. The box and whisker plot is generated from 4 replicate experiments. Statistical analyses by Tukey’s multiple comparisons test were performed using Prism software (GraphPad Software Inc.).

Article Snippet: To examine the biological relevance of this mechanism, we measured RHBDL2 activity and EGFR shedding in primary Normal Human Adult Epidermal Keratinocytes (NHEK-Ad) and hTERT-immortalized keratinocytes Ker-CT (ATCC CRL-4048), more stably accessible alternatives to primary keratinocytes.

Techniques: Western Blot, Migration, Microscopy, Sequencing, Plasmid Preparation, Transfection, Time-lapse Microscopy, Whisker Assay, shRNA, Mutagenesis, Over Expression, Control, Knockdown, Software, Alamar Blue Assay, Fluorescence, Activity Assay, Generated

Journal: bioRxiv

Article Title: Rhomboid protease RHBDL2 is a calcium-activated suppressor of EGFR signalling in keratinocytes

doi: 10.64898/2026.03.19.712941

Figure Lengend Snippet: A. RHBDL2 mRNA comparison by qPCR in the Ker-CT and HaCaT cells. Gene expression was normalized to GAPDH and the level of RHBDL2 expression in HaCaT cells was used to normalize RHBDL2 expression in all other cell lines, also in panel F. The mRNA analysis was done from three biological replicates, each in three technical replicates. Error bars show standard deviation. B. and C. Immunoblotting of conditioned medium and lysate after 4 h incubation with 5 mM calcium and 1 µM ionomycin detecting shedding of EGFR in primary NHEK-Ad and immortalized keratinocytes Ker-CT that is inhibited by a RHBDL2 ketoamide inhibitor compound 11 (50 µM) confirming RHBDL2 dependency. Tubulin is was used as a loading control. D. Immunoblotting of conditioned medium after 4 h incubation of Ker-CT keratinocytes with 100 µM PLCγ activator m-3M3FBS in the absence and presence of RHBDL2 inhibitor compound 11 (50 µM). E. Immunoblotting of conditioned medium after 4 h incubation of Ker-CT with thapsigargin (2 µM), bradykinin (10 µM) or 5 mM calcium as a positive control that induces RHBDL2 dependent EGFR shedding. F. RHBDL2 mRNA detection by qPCR in the N/TERT keratinocyte derived RHBDL2 knockout (R2ko) cell lines with comparison to the HaCaT and HaCaT RHBDL2 knockdown (R2kd) cells. The mRNA analysis was done from three biological replicates, each in three technical replicates. Error bars show standard deviation. G. Immunoblotting of conditioned medium and lysate of N/TERT keratinocytes to confirm RHBDL2 dependency of EGFR shedding in these cells and to validate the RHBDL2 KO (CB and CC) generated in N/TERT keratinocytes. Constitutive (48 h) or calcium (5 mM) stimulated (4 h) shedding of EGFR is inhibited by compound 11 (50 µM) in the WT, confirming its RHBDL2 dependence.

Article Snippet: To examine the biological relevance of this mechanism, we measured RHBDL2 activity and EGFR shedding in primary Normal Human Adult Epidermal Keratinocytes (NHEK-Ad) and hTERT-immortalized keratinocytes Ker-CT (ATCC CRL-4048), more stably accessible alternatives to primary keratinocytes.

Techniques: Comparison, Gene Expression, Expressing, Standard Deviation, Western Blot, Incubation, Control, Positive Control, Derivative Assay, Knock-Out, Knockdown, Generated

Journal: bioRxiv

Article Title: Rhomboid protease RHBDL2 is a calcium-activated suppressor of EGFR signalling in keratinocytes

doi: 10.64898/2026.03.19.712941

Figure Lengend Snippet: A. Hematoxylin and eosin staining of human skin equivalent sections generated from wild type (WT) and RHBDL2 deficient (clones CB and CC) N/TERT keratinocytes. B. Quantification of qualitative assessment of morphology of human skin organotypic cultures is based on histological analysis. Detailed scoring of individual parameters in each layer of WT or RHBDL2 deficient clones was rated on a scale 1 to 18 where 18 denotes highest differentiation ( Fig. S6 ). Differentiation scores between groups were compared by one-way ANOVA; adjusted P-value: ****<0.0001, ***≤0.001, N=4 for each group.

Article Snippet: To examine the biological relevance of this mechanism, we measured RHBDL2 activity and EGFR shedding in primary Normal Human Adult Epidermal Keratinocytes (NHEK-Ad) and hTERT-immortalized keratinocytes Ker-CT (ATCC CRL-4048), more stably accessible alternatives to primary keratinocytes.

Techniques: Staining, Generated, Clone Assay

Journal: bioRxiv

Article Title: Rhomboid protease RHBDL2 is a calcium-activated suppressor of EGFR signalling in keratinocytes

doi: 10.64898/2026.03.19.712941

Figure Lengend Snippet:

Article Snippet: To examine the biological relevance of this mechanism, we measured RHBDL2 activity and EGFR shedding in primary Normal Human Adult Epidermal Keratinocytes (NHEK-Ad) and hTERT-immortalized keratinocytes Ker-CT (ATCC CRL-4048), more stably accessible alternatives to primary keratinocytes.

Techniques: Quantitative Proteomics, Membrane, Mass Spectrometry