primary cell lines normal human epidermal keratinocytes nhek  (PromoCell)

Journal: The Kaohsiung journal of medical sciences

Article Title: LncRNA UCA1 negatively regulates NF-kB activity in psoriatic keratinocytes through the miR125a-A20 axis.

doi: 10.1002/kjm2.12363

Figure Lengend Snippet: FIGURE 1 LncRNA UCA1 is down-regulated in psoriatic keratinocytes. (A) Real-time PCR analysis of lncRNA UCA1 level in normal controls and psoriatic skin samples from 20 patients with psoriasis. (B, C) the proliferation rates of HaCat, activated-HaCat, NHEK and activated-NHEK cells were measured by MTT assay. (D, E) the level of IL-6, IL-8, and IFN-γ produced by HaCat, activated-HaCat, NHEK, and activated-NHEK were detected by ELISA assay. (F, G) expression levels of lncRNA UCA1 in HaCat, activated-HaCat, NHEK and activated-NHEK cells were measured by real-time PCR analysis. Data were shown as mean ± SD. LncRNA, long noncoding RNAs. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Human immortalized keratinocyte cell line HaCat and normal human epidermal keratinocyte cell line NHEK were purchased from ATCC (Manassas, VA) and cultured in DMEM (SH30022.01, HyClone, South Logan, UT) supplemented with 10% fetal bovine serum (FBS, FB25015, Clark,Richmond, VA) and 1% streptomy- cin and penicillin (SV30010, Hyclone) at 37 C in humidified air of 5% CO2.

Techniques: Real-time Polymerase Chain Reaction, MTT Assay, Produced, Enzyme-linked Immunosorbent Assay, Expressing

Journal: The Kaohsiung journal of medical sciences

Article Title: LncRNA UCA1 negatively regulates NF-kB activity in psoriatic keratinocytes through the miR125a-A20 axis.

doi: 10.1002/kjm2.12363

Figure Lengend Snippet: FIGURE 2 NF-κB is activated in psoriatic keratinocytes. (A) NF-κB activity of patients with psoriasis, (B) activated-HaCat, and (C) activated- NHEK cells were measured by Elisa assay. Data were shown as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: Human immortalized keratinocyte cell line HaCat and normal human epidermal keratinocyte cell line NHEK were purchased from ATCC (Manassas, VA) and cultured in DMEM (SH30022.01, HyClone, South Logan, UT) supplemented with 10% fetal bovine serum (FBS, FB25015, Clark,Richmond, VA) and 1% streptomy- cin and penicillin (SV30010, Hyclone) at 37 C in humidified air of 5% CO2.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay

Journal: The Kaohsiung journal of medical sciences

Article Title: LncRNA UCA1 negatively regulates NF-kB activity in psoriatic keratinocytes through the miR125a-A20 axis.

doi: 10.1002/kjm2.12363

Figure Lengend Snippet: FIGURE 3 LncRNA UCA1 negatively regulates NF-κB activity in psoriatic keratinocytes. (A, B) sh-UCA1 or pcDNA-UCA1 was transfected into activated-HaCat (A) and activated-NHEK (B) cells. The transfection efficiency was determined by real-time PCR analysis. (C, D) NF-κB activity of lncRNA UCA1 knockdown or overexpressed activated-HaCat (C) and activated-NHEK (D) cells was detected by ELISA assay. (E, F) the levels of IL-6, IL-8 and IFN-γ secreted by lncRNA UCA1 knockdown or overexpressed activated-HaCat (E) and activated-NHEK (F) were detected by Elisa assay. LncRNA, long noncoding RNAs

Article Snippet: Human immortalized keratinocyte cell line HaCat and normal human epidermal keratinocyte cell line NHEK were purchased from ATCC (Manassas, VA) and cultured in DMEM (SH30022.01, HyClone, South Logan, UT) supplemented with 10% fetal bovine serum (FBS, FB25015, Clark,Richmond, VA) and 1% streptomy- cin and penicillin (SV30010, Hyclone) at 37 C in humidified air of 5% CO2.

Techniques: Activity Assay, Transfection, Real-time Polymerase Chain Reaction, Knockdown, Enzyme-linked Immunosorbent Assay

Journal: The Kaohsiung journal of medical sciences

Article Title: LncRNA UCA1 negatively regulates NF-kB activity in psoriatic keratinocytes through the miR125a-A20 axis.

doi: 10.1002/kjm2.12363

Figure Lengend Snippet: FIGURE 5 LncRNA UCA1 positively regulated the expression level of A20in psoriatic keratinocytes. The protein level of A20 in patients with psoriasis (A, D), lncRNA UCA1 knockdown (B, E) or overexpressed (C, F) activated-HaCat and activated-NHEK cells was detected by western blotting (D–F). Numbers indicated the ratio of the optical density of the A20 band to the β-Actin band were obtained from three independent assays by using ImageJ software. LncRNA, long noncoding RNAs

Article Snippet: Human immortalized keratinocyte cell line HaCat and normal human epidermal keratinocyte cell line NHEK were purchased from ATCC (Manassas, VA) and cultured in DMEM (SH30022.01, HyClone, South Logan, UT) supplemented with 10% fetal bovine serum (FBS, FB25015, Clark,Richmond, VA) and 1% streptomy- cin and penicillin (SV30010, Hyclone) at 37 C in humidified air of 5% CO2.

Techniques: Expressing, Knockdown, Western Blot, Software

Journal: The Kaohsiung journal of medical sciences

Article Title: LncRNA UCA1 negatively regulates NF-kB activity in psoriatic keratinocytes through the miR125a-A20 axis.

doi: 10.1002/kjm2.12363

Figure Lengend Snippet: FIGURE 4 LncRNA UCA1 inhibits the expression of miR125a in psoriatic keratinocytes. (A) the interaction region between lncRNA UCA1 andmiR125a was predicted by microRNA.org and miRBase. (B–D) MiR125aexpression levels of patients with psoriasis (B), and lncRNA UCA1 knockdown or overexpressed activated-HaCat (C) and activated-NHEK (D) cells were determined by real-time PCR analysis. Data were shown as mean ± SD. LncRNA, long noncoding RNAs. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: Human immortalized keratinocyte cell line HaCat and normal human epidermal keratinocyte cell line NHEK were purchased from ATCC (Manassas, VA) and cultured in DMEM (SH30022.01, HyClone, South Logan, UT) supplemented with 10% fetal bovine serum (FBS, FB25015, Clark,Richmond, VA) and 1% streptomy- cin and penicillin (SV30010, Hyclone) at 37 C in humidified air of 5% CO2.

Techniques: Expressing, Knockdown, Real-time Polymerase Chain Reaction

Journal: International journal of biological macromolecules

Article Title: Design and development of multibiocomponent hybrid alginate hydrogels and lipid nanodispersion as new materials for medical and cosmetic applications.

doi: 10.1016/j.ijbiomac.2024.134405

Figure Lengend Snippet: Fig. 22. Short-term cytotoxicity after 1 h (A) and 3 h (B) incubation with tested materials on NHEK cells.

Article Snippet: Normal human dermal fibroblast (NHDF) and normal human keratinocytes (NHEK) cell lines were purchased from PromoCell.

Techniques: Incubation

Journal: International journal of biological macromolecules

Article Title: Design and development of multibiocomponent hybrid alginate hydrogels and lipid nanodispersion as new materials for medical and cosmetic applications.

doi: 10.1016/j.ijbiomac.2024.134405

Figure Lengend Snippet: Fig. 23. Long-term cytotoxicity on NHDF (A) and NHEK (B) cells. Cells were pre-incubated with tested materials for 1 h and 3 h.

Article Snippet: Normal human dermal fibroblast (NHDF) and normal human keratinocytes (NHEK) cell lines were purchased from PromoCell.

Techniques: Incubation

Journal: Journal of Inflammation Research

Article Title: The Effect of Osthole on Transient Receptor Potential Channels: A Possible Alternative Therapy for Atopic Dermatitis

doi: 10.2147/JIR.S425978

Figure Lengend Snippet: The level of IL-4 ( A and B ) and IL-13 ( C and D ) after incubation with histamine (Hist; 100 µg/mL), lipopolysaccharides (LPS; 2 µg/mL) alone and in mixtures with osthole (OST; 0.125 and 0.5 mg/mL) and clobetasol propionate (CP; 0.5 mg/mL) in Normal Human Epithelial Keratinocytes (NHEK; ( A and C )) and prepared model of 3D skin (3D skin; ( B and D )). The horizontal line shows the mean and the bars show the standard deviation. Statistically significant differences (Two-way ANOVA with Tukey’s multiple comparisons test) compared to control (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001) and to cells treated with histamine or LPS ( # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001) are marked.

Article Snippet: In brief, the Normal Human Epidermal Keratinocyte (NHEK) cell line was purchased from PromoCell GmbH (Heidelberg, Germany, cat. no. C-12005) and cultured in T-75 flasks in keratinocyte medium (Keratinocyte Growth Medium 2 ready to use, PromoCell, Heidelberg, Germany, cat. No. C-20011).

Techniques: Incubation, Standard Deviation, Control

Journal: Journal of Inflammation Research

Article Title: The Effect of Osthole on Transient Receptor Potential Channels: A Possible Alternative Therapy for Atopic Dermatitis

doi: 10.2147/JIR.S425978

Figure Lengend Snippet: Histamine receptor 4 ( A and B ) and IL-4 receptor-α ( C and D ) gene expression level after incubation with histamine (Hist; 100 µg/mL), lipopolysaccharides (LPS; 2 µg/mL) alone and in mixtures with osthole (OST; 0.125 and 0.5 mg/mL) and clobetasol propionate (CP; 0.5 mg/mL) in Normal Human Epithelial Keratinocytes (NHEK; ( A and C )) and prepared model of 3D skin (3D skin; ( B and D )). The horizontal line shows the mean and the bars show the standard deviation. Statistically significant differences (Two-way ANOVA with Tukey’s multiple comparisons test) compared to control (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001) and to cells treated with histamine or LPS (# p < 0.05, ### p < 0.001, #### p < 0.0001) are marked.

Article Snippet: In brief, the Normal Human Epidermal Keratinocyte (NHEK) cell line was purchased from PromoCell GmbH (Heidelberg, Germany, cat. no. C-12005) and cultured in T-75 flasks in keratinocyte medium (Keratinocyte Growth Medium 2 ready to use, PromoCell, Heidelberg, Germany, cat. No. C-20011).

Techniques: Gene Expression, Incubation, Standard Deviation, Control

Journal: Journal of Inflammation Research

Article Title: The Effect of Osthole on Transient Receptor Potential Channels: A Possible Alternative Therapy for Atopic Dermatitis

doi: 10.2147/JIR.S425978

Figure Lengend Snippet: TRPV1 ( A and B ), TRPV4 ( C and D ) and TRPM8 ( E and F ) gene expression level after incubation with histamine (Hist; 100 µg/mL), lipopolysaccharides (LPS; 2 µg/mL) alone and in mixtures with osthole (OST; 0.125 and 0.5 mg/mL) and clobetasol propionate (CP; 0.5 mg/mL) in Normal Human Epithelial Keratinocytes (NHEK; ( A, C and E )) and prepared model of 3D skin (3D skin; ( B, D and F )). The horizontal line shows the mean and the bars show the standard deviation. Statistically significant differences (Two-way ANOVA with Tukey’s multiple comparisons test) compared to control (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001) and to cells treated with histamine or LPS ( # p < 0.05, ### p < 0.001, #### p < 0.0001) are marked.

Article Snippet: In brief, the Normal Human Epidermal Keratinocyte (NHEK) cell line was purchased from PromoCell GmbH (Heidelberg, Germany, cat. no. C-12005) and cultured in T-75 flasks in keratinocyte medium (Keratinocyte Growth Medium 2 ready to use, PromoCell, Heidelberg, Germany, cat. No. C-20011).

Techniques: Gene Expression, Incubation, Standard Deviation, Control

Journal: Aging (Albany NY)

Article Title: Curcumol may alleviate psoriasis-like inflammation by inhibiting keratinocyte proliferation and inflammatory gene expression via JAK1/STAT3 signaling

doi: 10.18632/aging.203287

Figure Lengend Snippet: Curcumol inhibits proliferation and cell cycle progression and promote apoptosis in stimulated human keratinocytes. ( A ) CCK-8 cell viability assay in NHEK cells treated with curcumol (0, 5, 10, 20, 40 or 80 μM) for 24 h. ( B ) CCK-8 cell viability assay in NHEK cells pre-incubated with curcumol (10 or 20 μM) for 24 h, and stimulated with 10 ng/mL of M5 mix for 24 h. ( C ) Relative fluorescence levels were quantified by EdU and DAPI staining. ( D , E ) Flow cytometer assay was used to analyze cell apoptosis. ( F ) Cell cycle distribution was measured by flow cytometry. ( G – I ) Western blot was performed to measure the expressions of p27 Kip1 and CDK4 in NHEK cells. **P<0.01 compared with control group; # P<0.05, ## P<0.01 compared with 10 ng/mL_M5 group.

Article Snippet: Normal human epidermal keratinocytes (NHEK) cell line was purchased from ScienCell Research Laboratories (Carlsbad, CA, USA).

Techniques: CCK-8 Assay, Viability Assay, Incubation, Fluorescence, Staining, Flow Cytometry, Western Blot, Control

Journal: Aging (Albany NY)

Article Title: Curcumol may alleviate psoriasis-like inflammation by inhibiting keratinocyte proliferation and inflammatory gene expression via JAK1/STAT3 signaling

doi: 10.18632/aging.203287

Figure Lengend Snippet: Curcumol suppresses inflammatory gene expression in M5-treated NHEK cells. NHEK cells were preincubated with curcumol (10 or 20 μM) for 24 h, stimulated with M5 (10 ng/mL) for 24 h, and gene expression of IL-6 ( A ), IL-1β ( B ), CXCL1 ( C ), CXCL2 ( D ), LL37 ( E ), β-defensin-2 ( F ), S100A7 ( G ), and S100A8 ( H ) was analyzed by RT-qPCR. **P<0.01 compared with control group; # P<0.05, ## P<0.01 compared with 10 ng/mL_M5 group.

Article Snippet: Normal human epidermal keratinocytes (NHEK) cell line was purchased from ScienCell Research Laboratories (Carlsbad, CA, USA).

Techniques: Gene Expression, Quantitative RT-PCR, Control

Journal: Aging (Albany NY)

Article Title: Curcumol may alleviate psoriasis-like inflammation by inhibiting keratinocyte proliferation and inflammatory gene expression via JAK1/STAT3 signaling

doi: 10.18632/aging.203287

Figure Lengend Snippet: Curcumol attenuates M5-induced oxidative stress damage in NHEK cells. NHEK cells were treated with curcumol (10 or 20 μM) for 24 h, and then stimulated with M5 (10 ng/mL) for 24 h. ( A ) ROS production in NHEK cells was detected by flow cytometry. ( B – E ) ELISA was used to detect the levels of SOD ( B ), MDA ( C ), GSH ( D ), and CAT ( E ) in the supernatants of NHEK cells. **P<0.01 compared with control group; # P<0.05, ## P<0.01 compared with 10 ng/mL_M5 group.

Article Snippet: Normal human epidermal keratinocytes (NHEK) cell line was purchased from ScienCell Research Laboratories (Carlsbad, CA, USA).

Techniques: Flow Cytometry, Enzyme-linked Immunosorbent Assay, Control

Journal: Aging (Albany NY)

Article Title: Curcumol may alleviate psoriasis-like inflammation by inhibiting keratinocyte proliferation and inflammatory gene expression via JAK1/STAT3 signaling

doi: 10.18632/aging.203287

Figure Lengend Snippet: Curcumol promotes differentiation in M5-stimulated NHEK cells. NHEK cells were treated with curcumol (10 or 20 μM) for 24 h, and then stimulated with M5 (10 ng/mL) for 24 h. ( A ) Expression of keratin 1, keratin 5, keratin 10, filaggrin and loricrin in NHEK cells was analyzed by western blotting. ( B – F ) The relative expression of keratin 1 ( B ), keratin 5 ( C ), keratin 10 ( D ), filaggrin ( E ), and loricrin ( F ) quantified via normalization to β-actin. **P<0.01 compared with control group; # P<0.05, ## P<0.01 compared with 10 ng/mL_M5 group.

Article Snippet: Normal human epidermal keratinocytes (NHEK) cell line was purchased from ScienCell Research Laboratories (Carlsbad, CA, USA).

Techniques: Expressing, Western Blot, Control

Journal: Aging (Albany NY)

Article Title: Curcumol may alleviate psoriasis-like inflammation by inhibiting keratinocyte proliferation and inflammatory gene expression via JAK1/STAT3 signaling

doi: 10.18632/aging.203287

Figure Lengend Snippet: Curcumol inhibits proliferation of M5-treated NHEK cells via downregulation of p-STAT3/cyclin D2 axis. NHEK cells were treated with curcumol (10 or 20 μM) for 24 h, and then stimulated with M5 (10 ng/mL) for 24 h. ( A ) The expression level of p-STAT3 in NHEK cells was detected by western blotting, and normalized to STAT3. ( B ) The nuclear levels of p-STAT3 were analyzed by western blotting, and normalized to Histone H3. ( C ) Cyclin D2 mRNA levels were analyzed by RT-qPCR. ( D ) Cyclin D2 protein levels were analyzed by western blotting, and normalized to β-actin. **P<0.01 compared with control group; ## P<0.01 compared with 10 ng/mL_M5 group.

Article Snippet: Normal human epidermal keratinocytes (NHEK) cell line was purchased from ScienCell Research Laboratories (Carlsbad, CA, USA).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Control

Journal: Aging (Albany NY)

Article Title: Curcumol may alleviate psoriasis-like inflammation by inhibiting keratinocyte proliferation and inflammatory gene expression via JAK1/STAT3 signaling

doi: 10.18632/aging.203287

Figure Lengend Snippet: Curcumol inhibits proliferation of M5-treated NHEK cells via downregulation of JAK1/STAT3 signaling. ( A ) NHEK cells were treated with curcumol (10 or 20 μM; 24 h), and stimulated with M5 (10 ng/mL; 24 h). Levels of p-JAK1 were analyzed by western blotting, and normalized to JAK1. ( B ) NHEK cells were treated with 20 μM curcumol or/and 3μM ruxolitinib (RUX) for 24 h, and then stimulated with M5 (10 ng/mL) for 24 h. Levels of p-STAT3 were analyzed by western blotting, and normalized to STAT3. **P<0.01 compared with control group; # P<0.05, ## P<0.01 compared with M5 group; ^^P<0.01 compared with M5 + RUX group.

Article Snippet: Normal human epidermal keratinocytes (NHEK) cell line was purchased from ScienCell Research Laboratories (Carlsbad, CA, USA).

Techniques: Western Blot, Control