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: Current Research in Toxicology

Article Title: New approach methods for assessing indoor air toxicity

doi: 10.1016/j.crtox.2022.100090

Figure Lengend Snippet: The association between sampling site, reported health effects and the adverse effects detected in the biological methods.

Article Snippet: A genetically modified human keratinocyte cell line LuSens was kindly provided by BASF, Germany for research purposes.

Techniques: Sampling

Journal: Current Research in Toxicology

Article Title: New approach methods for assessing indoor air toxicity

doi: 10.1016/j.crtox.2022.100090

Figure Lengend Snippet: Description of biological and chemical analyses used to assess the quality of the indoor air samples (condensates).

Article Snippet: A genetically modified human keratinocyte cell line LuSens was kindly provided by BASF, Germany for research purposes.

Techniques: Activity Assay, Activation Assay, Purification, Binding Assay, Disruption, Concentration Assay

Journal: Current Research in Toxicology

Article Title: New approach methods for assessing indoor air toxicity

doi: 10.1016/j.crtox.2022.100090

Figure Lengend Snippet: Summary of the biological analyses of the indoor air samples. The three most sensitive assays to detect adverse effects in indoor air samples are bolded.

Article Snippet: A genetically modified human keratinocyte cell line LuSens was kindly provided by BASF, Germany for research purposes.

Techniques: Inhibition, Activity Assay, Activation Assay, Binding Assay, Disruption