LBR Antibody - #DF7356

FIGURE 2 RGR expression level and subcellular localization in human skin cells. (A) In normal human skin tissue, coimmunostaining of RGR (red) with the melanocyte marker melan-A (green) and keratinocyte marker P-CK (pink) is shown. Scale bar = 20 μm. The results showed positive staining of the RGR in the basal layer of the skin, large sudoriferous glands, hair follicles, and eccrine glands. Scale bar = 200 μm. (B) Representative expression of RGR and β-tubulin (loading control) in melanocytes, keratinocytes, and fibroblasts, as measured by the Western blot assay (anti-RGR antibody: a band of ∼32 kDa; β-tubulin antibody: a band of ∼55 kDa). The relative protein levels of RGR and β-tubulin were measured by ImageJ software (n = 3 independent experiments). (C) In cultured cells, the colocalization of the RGR (green) and melan-A (red) in melanocytes, colocalization of RGR (green) and CK-pan (red) in keratinocytes, and colocalization of RGR (green) and vimentin (red) in fibroblasts were detected by immunofluorescence double staining. Scale bar = 20 μm. (D) Localization and expression of the RGR (green) in human skin cells (melanocytes, keratinocytes, and fibroblasts) were observed by a confocal microscope. Scale bar = 20 μm. (E) IEM staining of the normal skin tissue. Representative protein A-gold probe represents RGR (RGR is mainly located in the nucleoplasm (yellow arrow) and mitochondria (red arrow) and scattered in the cytoplasm and other organelles (black arrow)). Scale bar = 1 μm. (F) Representative expression of RGR (green) in the membrane and nucleus of human keratinocytes was observed by confocal microscopy. Scale bar = 20 μm. (G) Representative differential expression of RGR, lamin B (nuclear control antibody), and GAPDH (loading control) in the human keratinocyte membrane, and the nucleus was analyzed by Western blotting. The relative protein levels were quantified using ImageJ software (n = 3 independent experiments). *p < 0.05, **p < 0.01, ***p < 0.001 vs the control group.