Human p63/TP73L Antibody

p63/TP73L in Human Breast. p63/TP73L was detected in immersion fixed paraffin-embedded sections of human breast using Goat Anti-Human p63/TP73L Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1916) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

p63/TP73L in SCC-25 Human Cell Line. p63/TP73L was detected in immersion fixed SCC-25 human tongue carcinoma cell line using Goat Anti-Human p63/TP73L Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1916) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (yellow; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Mouse p63/TP73L by Immunocytochemistry/Immunofluorescence Analysis of p73 and p63 co-expression in human and murine skin.(A) Scatter plot of TP63 versus TP73 RNA-seq expression [units = transcripts per million (TPM)] by human tissue type (n = 37) from the Human Protein Atlas (172 total samples) [30]. Mean expression (TPM + 0.1) for each tissue is plotted on a log2 scale with a LOESS smooth local regression line (gray). Correlation between TP63 and TP73 was quantified using Spearman’s rank correlation coefficient (rs). (B) Representative micrographs of H&E and immunofluorescence (IF) staining on serial human (top) and mouse (bottom) skin sections; DAPI (blue), p63 alpha (green), and p73 (red). Regions of the skin in micrographs are labeled as: interfollicular epidermis (IFE), hair follicle (HF), outer root sheath (ORS), HF bulge (Bu), hair bulb (HB), sebaceous gland (SG), hair shaft (HS), and arrector pili muscle (APM). Scale bars represent 200 μm for human and 50 μm for murine tissue. See also S1 and S2 Figs. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31216312), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human p63/TP73L by Western Blot Analysis of delta Np73 in epidermal programming using an induced basal keratinocyte (iKC) model system.(A) Immunoblot of KLF4, p63 alpha, and p73 protein expression in neonatal human dermal fibroblast (HDFn) cells infected with lentivirus encoding delta Np73 isoforms ( delta Np73 alpha and delta Np73 beta ) or empty vector control in combination with KLF4 and delta Np63 alpha. Cells were grown for 3 days and protein was harvested for immunoblot analysis. (B) Bar graphs of RNA expression for the indicated keratinocyte genes in HDFn cells infected in (A). Cells were grown for 3 days and RNA was harvested for qRT-PCR analysis. Expression data are represented as the fold increase relative to control. The mean of three replicates is shown with error bars representing SEM. *p-value < 0.05, **p-value < 0.01, ***p-value < 0.001. (C) Principal component analysis (PCA) plot of RNA-seq from HDFn cells infected with lentivirus encoding delta Np73 beta or empty vector control in combination with KLF4 and delta Np63 alpha. Cells were grown for 6 days and RNA was harvested for RNA-seq analysis. The percentage of variance contributed by each PC is listed in parentheses. (D and E) Tables listing the enriched Genome Ontology (GO) categories and pathways among the top 250 genes contributing to PC1 from (C). (F) Heatmap with the expression of a set of 44 genes that underlie the enrichment of GO categories from (D). Genes are annotated based on known roles in iKC-related processes (gray box) and the presence of a p63/p73 ChIP-seq peak within 50 kb of its TSS in multiple basal cell types (brown box). See also S5 and S6 Figs and S2–S9 Tables. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31216312), licensed under a CC-BY license. Not internally tested by R&D Systems.