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
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