UCHL1/PGP 9.5 Dynamic in Neuro-Immune-Cutaneous Milieu: Focusing on Axonal Nerve Terminals and Epidermal Keratinocytes in Psoriatic Itch

Figure 1

Expression of UCHL1 gene and PGP 9.5 protein in skin punch biopsies, HaCaT keratinocyte cell line, and brain reference material. Confocal scanning microcopy using HaCaT keratinocyte cell line presents images of cytoplasmatic expression of PGP 9.5 protein (a1). Furthermore, omission of primary antibody against human PGP 9.5 (a2) as well as IgG-AF488 isotype control does not result in any specific signal (a3). Scale bars represent 20μm. UCHL1 expression at mRNA level was observed in HaCaT cell line, normal skin, and was confirmed in brain reference material. Forward and reverse primers amplificate 95nt sequence at the border of exons 3 and 4 of UCHL1 gene detected using FAM-labeled UPL 27 probe. ACTB was used as housekeeping gene using VIC-labeled probe and amplificate 121nt fragment. All amplified material was visualized using gel electrophoresis (b). Protein expression was verified using Western-blot with the same mouse anti-human PGP 9.5 antibody and gave specific 27 kDa band in the skin, brain, and HaCaT cell line, respectively (c).