Fig. 2: UVA increases OPN3 expression in human epidermal keratinocytes.
From: Opsin 3 mediates UVA-induced keratinocyte supranuclear melanin cap formation
a, b RT-qPCR analysis of OPNs gene expression levels in HaCaT and HEK. OPNs mRNA levels were normalized to GAPDH levels (n = 3 independent experiments). Statistical significance was determined by one-ANOVA with post-test. ****P < 0.0001. c WB analysis of OPN3 protein expression in HaCaT and HEK. d, e Representative fluorescence confocal images of OPN3 protein expression in HaCaT (left) and HEK (right). OPN3 stained with anti-human OPN3 polyclonal antibody (green), the nucleus stained with DAPI (blue), and the keratinocytes marker Pan-Cytokeratin (Pan-CK) (Cy3, red). Scale bars = 20 µm. Immunofluorescence double staining also appeared positive OPN3 staining on the membrane and cytoplasmic of HaCaT and HEK. f HaCaT were irradiated with UVA (3 J cm−2, 5 J cm−2, 10 J cm−2, 15 J cm−2, 20 J cm−2), and OPN3 protein expression levels were determined by WB analysis. WB analyses were normalized using β-tubulin as a loading control, and the relative protein level was quantified using Quantity One software. n = 3 independent experiments. Statistical significance was determined by one-ANOVA with post-test. *P < 0.05; **P < 0.01. g HaCaT were irradiated with 3 J cm−2 UVA, and OPN3 mRNA levels were determined by RT-qPCR analysis. OPN3 mRNA levels were normalized to GAPDH levels (n = 3 independent experiments). Statistical significance was determined by t-test analysis. **P < 0.01. h HEK were irradiated with 3 J cm−2 UVA, and OPN3 protein expression levels were determined by WB analysis. WB analyses were normalized using β-tubulin as a loading control, and the relative protein level was quantified using Quantity One software. n = 3 independent experiments. Statistical significance was determined by t-test analysis. * P < 0.05; **P < 0.01. i HEK were irradiated with 3 J cm−2 UVA, and OPN3 mRNA levels were determined by RT-qPCR analysis. OPN3 mRNA levels were normalized to GAPDH levels. n = 3 independent experiments. Statistical significance was determined by t-test analysis. **P < 0.01.
