Journal: International Journal of Molecular Sciences

Article Title: Loss of BAX by miR-365 Promotes Cutaneous Squamous Cell Carcinoma Progression by Suppressing Apoptosis

doi: 10.3390/ijms18061157

Figure Lengend Snippet: BAX is decreased in cutaneous squamous cell carcinoma (CSCC) primary tumors and cell lines. ( A ) BAX and miR-365 expression was detected in the CSCC cell lines A431, HSC-5, and HaCaT miR-365 compared with HaCaT keratinocytes in both mRNA and protein levels. The qPCR results were evaluated by normalizing to U6 snRNA (for miR-365) or GAPDH (for BAX). In Western blot, GAPDH was detected for using as loading control; ( B ) BAX and miR-365 expression were detected in normal tissues (N1–N4) and CSCC primary tumors (C1–C6) in both mRNA and protein levels; ( C ) IHC detection of BAX on paraffin sections of CSCC tumors and normal skin specimens. Positive signals were shown in brown staining (magnification, 400×), scale bars, 50 µm. The percentage of positive staining was marked for each group. ** p < 0.01, *** p < 0.001.

Article Snippet: The CSCC lines A431, HSC-1 (HonSun Biological Co., Ltd., Shanghai, China), the human benign epidermal keratinocyte cell line HaCaT (China Center for Type Culture Collection, Wuhan, China), and the HaCaT miR-365 cell strain stably overexpressing miR-365 constructed in our previous study [ ] were cultured using DMEM (Dulbecco’s modified Eagle medium) with 10% fetal bovine serum and maintain at 37 °C with 5% CO 2 in a humidified environment.

Techniques: Expressing, Western Blot, Control, Staining

Journal: Frontiers in Immunology

Article Title: Antagonizing Retinoic Acid-Related-Orphan Receptor Gamma Activity Blocks the T Helper 17/Interleukin-17 Pathway Leading to Attenuated Pro-inflammatory Human Keratinocyte and Skin Responses

doi: 10.3389/fimmu.2019.00577

Figure Lengend Snippet: Cpd A attenuates IL-17-induced secondary responses in keratinocytes. (A) HaCaT keratinocytes were cultured for 24 h with supernatants derived from Cpd A treated Th17 cells either at a biologically inactive (0.13 nM) or at a pharmacologically active concentration (10 μM). To determine the effect of IL-17 in the conditioned supernatants on the HaCaT cell activation, half of the supernatants were pre-incubated with the IL-17 selective, neutralizing antibody Secukinumab (0.75 μg/ml). Alternatively, cells were left unstimulated or were exposed to Th17 cytokines that were used in the Th17 polarization assays. Keratinocytes were subjected to qRT-PCR analysis for determination of downstream IL-17-response genes, including NFKBIZ, DEFB4A, CCL20 , and IL36G . (B,C) Defined IL-17A and TNFα concentrations which were measured in the Th17 cell supernatants originating from low-Cpd A-containing samples (IL-17: 0.6 ng/ml; TNFα 4 ng/ml) and from high-Cpd A-containing (IL-17: 0.15 ng/ml; TNFα: 4 ng/ml) or Secukinumab-treated samples were spiked into HaCaT cells (B) or alternatively into NHEK cell cultures originating from two donors (C) . To determine the impact of IL-17 in this cellular system, IL-17 was neutralized by addition of Secukinumab (0.75 μg/ml). Graphs are representative from three experiments containing triplicate readings. Significance between low-Cpd A-containing supernatants and between high-Cpd A and Secukinumab-containing samples was determined by ANOVA followed by Dunnett's test (*** p < 0.005; **** p < 0.0001). Error bars represent the SD.

Article Snippet: The HaCaT keratinocyte cell line was purchased from AddexBio and was maintained in RPMI 1640 medium supplemented with Glutamax, Penicillin/Streptomycin (all from Gibco) and 10% FCS (PAA).

Techniques: Cell Culture, Derivative Assay, Concentration Assay, Activation Assay, Incubation, Quantitative RT-PCR

Journal: Nanomaterials

Article Title: Gold Nanoparticles Prepared with Phyllanthus emblica Fruit Extract and Bifidobacterium animalis subsp. lactis Can Induce Apoptosis via Mitochondrial Impairment with Inhibition of Autophagy in the Human Gastric Carcinoma Cell Line AGS

doi: 10.3390/nano11051260

Figure Lengend Snippet: Cytotoxic effect of PEFE-AuNPs against normal and cancer cells. ( A ) The hemocompatibility of PEFE-Au NPs and PEFE. ( B , C ) The cytotoxic effect of PEFE-AuNPs, PEFE, and B. lactis against normal (HaCaT and NHDF) and cancer (AGS) cells. The cisplatin was used as positive control. ( D – F ) Microscopic images and colony formation of AGS cells treated with PEFE-AuNPs and PEFE. The cisplatin was used as positive control. *, p < 0.05, **, p < 0.01, ***, p < 0.001.

Article Snippet: Two types of normal cell lines, the human epidermal keratinocyte cell line HaCaT and human dermal fibroblast cell line NHDF, were purchased from KCLB (Seoul, Korea).

Techniques: Positive Control

Journal: Marine Drugs

Article Title: Fucoidan Isolated from Sargassum confusum Suppresses Inflammatory Responses and Oxidative Stress in TNF-α/IFN-γ- Stimulated HaCaT Keratinocytes by Activating Nrf2/HO-1 Signaling Pathway

doi: 10.3390/md20020117

Figure Lengend Snippet: Protective effect of SCF in TNF-α/IFN-γ-stimulated HaCaT keratinocytes. ( A ) Time-dependent cell viability, ( B ) intracellular ROS generation, analysis of ROS level with and without TNF-α/IFN-γ stimulation by ( C ) fluorescence microscopy, and ( D ) flow cytometer. For ROS assays, cells were treated with DCF-DA. The results represent data from three independent experiments ( n = 3), and values are indicated as the means ± SE. Error bars with different letters are significantly different ( p ˂ 0.05).

Article Snippet: HaCaT keratinocytes purchased from the Korean Cell Line Bank (Seoul, Korea) were cultured at 37 °C in an incubator with a humidified environment of 5% CO 2 in DMEM medium supplemented with 10% FBS and 1% penicillin/streptomycin mixture.

Techniques: Fluorescence, Microscopy, Flow Cytometry

Journal: Marine Drugs

Article Title: Fucoidan Isolated from Sargassum confusum Suppresses Inflammatory Responses and Oxidative Stress in TNF-α/IFN-γ- Stimulated HaCaT Keratinocytes by Activating Nrf2/HO-1 Signaling Pathway

doi: 10.3390/md20020117

Figure Lengend Snippet: Inhibitory effect of SCF on mRNA expression of inflammatory cytokines and chemokines. Effect of SCF against the mRNA expression of ( A ) epithelial and epidermal innate cytokines, ( B ) inflammatory cytokines, ( C ) chemokines, ( D ) densitometry analysis, and ( E ) ELISA analysis of cytokines and chemokines production in TNF-α/IFN-γ-stimulated HaCaT keratinocytes. The reproducibility of the results was confirmed by three independent determinant determinations ( n = 3, mean ± SE). Error bars with different letters show a significant difference ( p ˂ 0.05).

Article Snippet: HaCaT keratinocytes purchased from the Korean Cell Line Bank (Seoul, Korea) were cultured at 37 °C in an incubator with a humidified environment of 5% CO 2 in DMEM medium supplemented with 10% FBS and 1% penicillin/streptomycin mixture.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Marine Drugs

Article Title: Fucoidan Isolated from Sargassum confusum Suppresses Inflammatory Responses and Oxidative Stress in TNF-α/IFN-γ- Stimulated HaCaT Keratinocytes by Activating Nrf2/HO-1 Signaling Pathway

doi: 10.3390/md20020117

Figure Lengend Snippet: Inhibitory effect of SCF on MAPK and NF-κB molecular mediators in TNF-α/IFN-γ-stimulated HaCaT keratinocytes. ( A ) Western blot analysis of key MAPK mediators, ( B ) phosphorylation and nuclear translocation of NF-κB molecular mediators. ( C ) Immunofluorescence analysis of NF-κB p65 nuclear translocation. All experiments were performed in triplicate ( n = 3), and values are indicated as the means ± SE. Error bars with different letters show a significant difference ( p ˂ 0.05).

Article Snippet: HaCaT keratinocytes purchased from the Korean Cell Line Bank (Seoul, Korea) were cultured at 37 °C in an incubator with a humidified environment of 5% CO 2 in DMEM medium supplemented with 10% FBS and 1% penicillin/streptomycin mixture.

Techniques: Western Blot, Phospho-proteomics, Translocation Assay, Immunofluorescence

Journal: Marine Drugs

Article Title: Fucoidan Isolated from Sargassum confusum Suppresses Inflammatory Responses and Oxidative Stress in TNF-α/IFN-γ- Stimulated HaCaT Keratinocytes by Activating Nrf2/HO-1 Signaling Pathway

doi: 10.3390/md20020117

Figure Lengend Snippet: Cytoprotective effect of SCF on the activation of Nrf2/HO-1 signaling pathway. ( A ) Dose-dependent response of the SCF on activation of HO-1 and NQO1, and ( B ) nuclear-translocated Nrf2 in TNF-α/IFN-γ-stimulated HaCaT keratinocytes by Western blot analysis. Data are represented as mean ± SE of three independent determinations ( n = 3). Mean values with different letters are significantly different ( p ˂ 0.05).

Article Snippet: HaCaT keratinocytes purchased from the Korean Cell Line Bank (Seoul, Korea) were cultured at 37 °C in an incubator with a humidified environment of 5% CO 2 in DMEM medium supplemented with 10% FBS and 1% penicillin/streptomycin mixture.

Techniques: Activation Assay, Western Blot

Journal: Marine Drugs

Article Title: Fucoidan Isolated from Sargassum confusum Suppresses Inflammatory Responses and Oxidative Stress in TNF-α/IFN-γ- Stimulated HaCaT Keratinocytes by Activating Nrf2/HO-1 Signaling Pathway

doi: 10.3390/md20020117

Figure Lengend Snippet: Effect of HO-1 inhibition by ZnPP on SCF treated ( A ) time-dependent cell viability, ( B ) intracellular ROS production, and ( C ) analysis of ROS levels by fluorescence microscopy in TNF-α/IFN-γ-stimulated HaCaT keratinocytes. The results represent data from three independent experiments ( n = 3), and values are indicated as the means ± SE. Error bars with different letters show a significant difference ( p ˂ 0.05).

Article Snippet: HaCaT keratinocytes purchased from the Korean Cell Line Bank (Seoul, Korea) were cultured at 37 °C in an incubator with a humidified environment of 5% CO 2 in DMEM medium supplemented with 10% FBS and 1% penicillin/streptomycin mixture.

Techniques: Inhibition, Fluorescence, Microscopy

Journal: Scientific Reports

Article Title: Low-intensity pulsed ultrasound induces multifaced alterations in chromosome segregation, cytoskeletal filaments and cell junctions

doi: 10.1038/s41598-025-88569-1

Figure Lengend Snippet: Micronucleus test with CREST staining. Representative images of a binucleated cell with CREST-negative micronucleus (MN-, A ) and binucleated cell with CREST-positive micronucleus (MN+, B ); kinetochores are stained in green, DNA in blue; Scale bar: 5 μm. Frequencies of CREST-positive (MN+), CREST-negative (MN-) and total micronuclei (MN) per binucleated cell (BNC) in HaCaT ( C ) and SK-MEL-28 ( D ) cells. Bars represent mean ± SD. Points represent values of each sample. Asterisks represent significant differences with unsonicated samples (* p < 0.05; ** p < 0.01).

Article Snippet: The immortalized human keratinocyte cell line, HaCaT, and the human melanoma cell line SK-MEL − 28 were obtained from Interlab Cell Line Collection (ICLC, National Cancer Research Institute, Genoa, Italy).

Techniques: Staining

Journal: Scientific Reports

Article Title: Low-intensity pulsed ultrasound induces multifaced alterations in chromosome segregation, cytoskeletal filaments and cell junctions

doi: 10.1038/s41598-025-88569-1

Figure Lengend Snippet: Effects of LIPUS exposure on F-actin filaments and cell adhesion. Confocal fluorescence of Phalloidin immunolocalization (green) in HaCaT ( A ) and SK-MEL-28 cells ( B ) treated for 1 h with 85 mW/cm 2 US (right) compared to control (left); scale bar 10 μm. C FE-SEM images of HaCaT cells: non-treated cells (left), exposed to LIPUS (65 mW/cm 2 , middle), and positive-treated control (160 mW/cm 2 , right). Scale bars: main images: 10 μm, bottom right images 1 μm. D FE-SEM images of HaCaT cell fractures following US irradiation: control (left), 65 mW/cm 2 (middle), 160 mW/cm 2 (right). Scale bar: 10 μm. Bottom graphs: profilometric analysis highlighting the breaks in the treated cells (65 mW/cm 2 , middle; 160 mW/cm 2 , right).

Article Snippet: The immortalized human keratinocyte cell line, HaCaT, and the human melanoma cell line SK-MEL − 28 were obtained from Interlab Cell Line Collection (ICLC, National Cancer Research Institute, Genoa, Italy).

Techniques: Fluorescence, Control, Irradiation

Journal: Scientific Reports

Article Title: Low-intensity pulsed ultrasound induces multifaced alterations in chromosome segregation, cytoskeletal filaments and cell junctions

doi: 10.1038/s41598-025-88569-1

Figure Lengend Snippet: Adherens junctions and desmosomes are modulated by LIPUS. A β-catenin and E-cadherin protein levels obtained by western blot analysis of cells following 85mW/cm 2 LIPUS irradiation. B and C Densitometric analysis of β-catenin and E-cadherin levels. D Western blot of Desmosome levels in LIPUS exposed HaCaT and SK-MEL-28 cells. β-Actin was used for protein levels normalization. One representative blot is shown for each antigen, data are presented as a mean ± SD ( n ≥ 3, Student’s t-test * p < 0.05; *** p < 0.001). E Immunofluorescence analysis of Desmosome in HaCaT cells. Magnification 100X; scale bar 10 μm. Full-length blots are shown in ESM (Section S4, Figure S6).

Article Snippet: The immortalized human keratinocyte cell line, HaCaT, and the human melanoma cell line SK-MEL − 28 were obtained from Interlab Cell Line Collection (ICLC, National Cancer Research Institute, Genoa, Italy).

Techniques: Western Blot, Irradiation, Immunofluorescence

Journal: Scientific Reports

Article Title: Low-intensity pulsed ultrasound induces multifaced alterations in chromosome segregation, cytoskeletal filaments and cell junctions

doi: 10.1038/s41598-025-88569-1

Figure Lengend Snippet: Analysis of cytokeratin levels and distribution upon LIPUS exposure ( A ) Western blot (upper) and densitometric analysis (lower) of cytokeratin in HaCaT cells. β-Actin was used as loading control. Immunostaining for cytokeratin in HaCat ( B ) and SK-MEL-28 ( D ) cells and the corresponding densitometric analysis ( C and E ). Both cell lines are shown after treatment with 85 mW/cm 2 LIPUS. One representative blot/image is reported for each antigen, data are presented as a mean ± SD ( n ≥ 3, Student’s t-test * p < 0.05; *** p < 0.001). Magnification 100X; scale bar 10 μm. Full-length blot is shown in ESM (Section S4, Figure S7).

Article Snippet: The immortalized human keratinocyte cell line, HaCaT, and the human melanoma cell line SK-MEL − 28 were obtained from Interlab Cell Line Collection (ICLC, National Cancer Research Institute, Genoa, Italy).

Techniques: Western Blot, Control, Immunostaining

Journal: Scientific Reports

Article Title: Low-intensity pulsed ultrasound induces multifaced alterations in chromosome segregation, cytoskeletal filaments and cell junctions

doi: 10.1038/s41598-025-88569-1

Figure Lengend Snippet: Percentages of binucleated cells. A HaCaT cells; B SK-MEL-28 cells. Bars represent mean ± SD. Points represent values of each sample. Asterisks represent significant differences with unsonicated samples (* p < 0.05; ** p < 0.01).

Article Snippet: The immortalized human keratinocyte cell line, HaCaT, and the human melanoma cell line SK-MEL − 28 were obtained from Interlab Cell Line Collection (ICLC, National Cancer Research Institute, Genoa, Italy).

Techniques:

Journal: Scientific Reports

Article Title: Low-intensity pulsed ultrasound induces multifaced alterations in chromosome segregation, cytoskeletal filaments and cell junctions

doi: 10.1038/s41598-025-88569-1

Figure Lengend Snippet: Mitotic indices. Mitotic index in HaCaT ( A ) and SK-MEL-28 cells ( B ). Ana-telophase / pro-metaphase ratio (A + T / P + M) in HaCaT ( C ) and SK-MEL-28 cells ( D ). Bars represent mean ± SD. Points represent values of each sample. Asterisks represent significant differences with unsonicated samples (* p < 0.05).

Article Snippet: The immortalized human keratinocyte cell line, HaCaT, and the human melanoma cell line SK-MEL − 28 were obtained from Interlab Cell Line Collection (ICLC, National Cancer Research Institute, Genoa, Italy).

Techniques:

Journal: Scientific Reports

Article Title: Low-intensity pulsed ultrasound induces multifaced alterations in chromosome segregation, cytoskeletal filaments and cell junctions

doi: 10.1038/s41598-025-88569-1

Figure Lengend Snippet: Effects of LIPUS treatment on Cyclin B1 and Caspase-3 levels. A The levels of Cyclin B1 were measured by western blot in HaCaT and SK-MEL-28 cells (upper) and quantified by densitometric analysis (lower). B Immunodetection of Cyclin B1 in HaCaT cells and C its corresponding signal analysis (magnification 100X; scale bar 10 μm) D Western blot analysis of Caspase-3 in HaCaT and SK-MEL-28 cells. The related densitometric analysis is reported below. E Immunofluorescence analysis of Caspase-3 and F its quantification in HaCaT cells (magnification 60X; scale bar 10 μm). All experiments were performed at the following setup: 85 mW/cm 2 for 1 h. One representative blot/image is reported for each antigen, data are presented as a mean ± SD ( n ≥ 3, Student’s t-test * p < 0.05; *** p < 0.001). Full-length blots are shown in ESM (Section S4, Figure S8).

Article Snippet: The immortalized human keratinocyte cell line, HaCaT, and the human melanoma cell line SK-MEL − 28 were obtained from Interlab Cell Line Collection (ICLC, National Cancer Research Institute, Genoa, Italy).

Techniques: Western Blot, Immunodetection, Immunofluorescence