recombinant human hbegf  (R&D Systems)

Journal: Experimental dermatology

Article Title: Pharmacological Impacts of Mucopolysacccharide Polyphosphates in the Epidermis Involves Inhibition of Amphiregulin-Mediated Signals in Keratinocytes.

doi: 10.1111/exd.70000

Figure Lengend Snippet: FIGURE 3 | MPS induced perturbations similar to those resulting from AREG inhibition. (a) A bar chart showing the TPM levels of seven known EGFR-ligands in untreated monolayer-cultured keratinocytes. Data represent the mean ± SD (n = 3). (b) A bar chart showing the first principal component value from PCA of bulk RNA-seq data from keratinocytes, subjected to various treatments for 24 h: 50 μg/mL of AREG antibody (AREG_Ab), 300 ng/ mL of AREG, 30 μg/mL of MPS, or a combination of AREG and MPS. Data represent the mean ± SD (n = 3). *p < 0.05 versus untreated control (Dunnett's multiple comparison test), #p < 0.05 versus AREG-treated (Aspin-Welch's t-test). (c) A heatmap showing the expression of 4150 DEGs from the monolayer culture experiment (Figure 1) in keratinocytes subjected to the aforementioned treatments. The colour scale represents the scaled expression of each gene. (d) Enrichment plots from GSEA of DEGs in keratinocytes treated with AREG antibody, compared to untreated controls. Top 200 upregulated or downregulated genes from the monolayer culture experiment (Figure 1) were utilised as gene sets. P denotes p value, NES denotes the normalised enrichment score. (e) Relative expression of representative genes in keratinocytes, as determined by real-time qPCR. These keratinocytes were treated with MPS at concentrations ranging from 3 to 300 μg/mL for 24 h, or untreated, in the presence of various EGFR-ligands. The concentrations of the EGFR- ligands were as follows: EGF at 3 ng/mL, HBEGF at 10 ng/mL, AREG at 300 ng/mL, and EPGN at 1000 ng/mL. Data represent the mean ± SD (n = 3). DEG, differentially expressed gene; GSEA, gene set enrichment analysis; MPS, mucopolysacccharide polyphosphate; TPM, transcripts per million.

Article Snippet: After HEKa cells had been seeded on cell culture plates at a density of 5.6 × 104 cells/cm2 and incubated for 24 h, then they were incubated with or without MPS (Maruho, Osaka, Japan), recombinant human amphiregulin (R&D systems, Minneapolis, MN, USA), human amphiregulin antibody (R&D systems), recombinant human EGF (R&D systems), recombinant human HBEGF (R&D systems), recombinant human EREG (R&D systems), in HuMedia- KB2 (Kurabo Industries) for 24 h at 37°C.

Techniques: Inhibition, Cell Culture, RNA Sequencing, Control, Comparison, Expressing