Expression of Opsin Molecule in Cultured Murine Melanocyte

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Expression of Opsin Molecule in Cultured Murine Melanocyte Yoko Miyashita, Tsuneo Moriya, Kouichi Asami Journal of Investigative Dermatology Symposium Proceedings Volume 6, Issue 1, Pages 54-57 (November 2001) DOI: 10.1046/j.0022-202x.2001.00018.x Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Agarose gel electrophoresis of the PCR products amplified with the primers for rhodopsin mRNA. The PCR analyses using cDNA of the murine melanocyte as the template. Lane 1, RM4/RM3; lane 2, RM1/RM3; lane 3, RM1/RM2; M, marker, 61°C, 37 cycles. The arrows indicate the PCR products (expected sizes; 183, 729, and 567 bp, respectively). Journal of Investigative Dermatology Symposium Proceedings 2001 6, 54-57DOI: (10.1046/j.0022-202x.2001.00018.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Western blot analysis of cultured murine melanocytes using polyclonal antibody against bovine-RH (1:5000) or normal serum. Lanes 1 and 2, bovine-RH (1:5000); lane 3, normal serum. Lane 1, bovine rhodopsin (2.7 µg protein); lanes 2 and 3 were each loaded with 10 µg of whole-cell protein extract from the melanocytes. Journal of Investigative Dermatology Symposium Proceedings 2001 6, 54-57DOI: (10.1046/j.0022-202x.2001.00018.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Immunofluoresence microscopy of cultured murine melanocyte using polyclonal antibody against rhodopsin. Melanocytes were treated with peptide-RH AB (1:500) (see Materials and Methods) (A) and without the antibody (B). Each triplet (a–c) of (A) and (B) represents the same field. The cells were visualized by FITC staining (a) and by Hoechst staining (b). Scale bars: 50 µm. Journal of Investigative Dermatology Symposium Proceedings 2001 6, 54-57DOI: (10.1046/j.0022-202x.2001.00018.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Agarose gel electrophoresis of the PCR products amplified with the primer for rhodopsin mRNA (RM1/RM3). (A) The RT-PCR analysis using total RNA of murine lymphoma cell line, EL4. Lane 1, EL4; lane 2, eye tissue; lane 3, Melan a2. (B) The RT-PCR analysis using total RNA of various murine tissues as well as that of the murine melanocytes. M, marker; lane 1, skin; lane 2, liver; lane 3, eye; lane 4, Melan a2; lane 5, without DNA. 61°C, 40 cycles. The arrows indicate the PCR products (expected sizes; 729 bp). Journal of Investigative Dermatology Symposium Proceedings 2001 6, 54-57DOI: (10.1046/j.0022-202x.2001.00018.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions