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Detection of Dystrophin Dp71 in Human Skeletal Muscle Using an Automated Capillary Western Assay System.
Kawaguchi, Tatsuya; Niba, Emma Tabe Eko; Rani, Abdul Qawee Mahyoob; Onishi, Yoshiyuki; Koizumi, Makoto; Awano, Hiroyuki; Matsumoto, Masaaki; Nagai, Masashi; Yoshida, Shinobu; Sakakibara, Sachiko; Maeda, Naoyuki; Sato, Osamu; Nishio, Hisahide; Matsuo, Masafumi.
Afiliação
  • Kawaguchi T; Discovery Science and Technology Department, Daiichi Sankyo RD Novare Co., Ltd., Edogawa, Tokyo 134-8634, Japan. kawaguchi.tatsuya.sg@rdn.daiichisankyo.co.jp.
  • Niba ETE; Department of Physical Therapy, Faculty of Rehabilitation, Kobe Gakuin University, Nishi, Kobe 651-2180, Japan. niba@med.kobe-u.ac.jp.
  • Rani AQM; Department of Community Medicine and Social Healthcare Sciences, Kobe University Graduate School of Medicine, Chuo, Kobe 650-0017, Japan. niba@med.kobe-u.ac.jp.
  • Onishi Y; Department of Physical Therapy, Faculty of Rehabilitation, Kobe Gakuin University, Nishi, Kobe 651-2180, Japan. rani@reha.kobegakuin.ac.jp.
  • Koizumi M; Modality Research Laboratories, Biologics Division, Daiichi Sankyo Co., Ltd., Shinagawa, Tokyo 140-8710, Japan. onishi.yoshiyuki.a2@daiichisankyo.co.jp.
  • Awano H; Modality Research Laboratories, Biologics Division, Daiichi Sankyo Co., Ltd., Shinagawa, Tokyo 140-8710, Japan. koizumi.makoto.h7@daiichisankyo.co.jp.
  • Matsumoto M; Department of Pediatrics, Kobe University Graduate School of Medicine, Chuo, Kobe 650-0017, Japan. awahiro@med.kobe-u.ac.jp.
  • Nagai M; Department of Pediatrics, Kobe University Graduate School of Medicine, Chuo, Kobe 650-0017, Japan. mmatsu@med.kobe-u.ac.jp.
  • Yoshida S; Department of Pediatrics, Kobe University Graduate School of Medicine, Chuo, Kobe 650-0017, Japan. natsu@med.kobe-u.ac.jp.
  • Sakakibara S; Discovery Science and Technology Department, Daiichi Sankyo RD Novare Co., Ltd., Edogawa, Tokyo 134-8634, Japan. yoshida.Shinobu.z3@rdn.daiichisankyo.co.jp.
  • Maeda N; Biomarker Department, Oncology Function, Daiichi Sankyo Co., Ltd., Shinagawa, Tokyo 140-8710, Japan. sakakibara.sachiko.bk@daiichisankyo.co.jp.
  • Sato O; Biomarker Department, Oncology Function, Daiichi Sankyo Co., Ltd., Shinagawa, Tokyo 140-8710, Japan. maeda.naoyuki.ct@daiichisankyo.co.jp.
  • Nishio H; R&D Planning & Management Department, Daiichi Sankyo Co., Ltd., Shinagawa, Tokyo 140-8710, Japan. sato.osamu.ua@daiichisankyo.co.jp.
  • Matsuo M; Department of Community Medicine and Social Healthcare Sciences, Kobe University Graduate School of Medicine, Chuo, Kobe 650-0017, Japan. nishio@med.kobe-u.ac.jp.
Int J Mol Sci ; 19(6)2018 May 23.
Article em En | MEDLINE | ID: mdl-29789502
ABSTRACT

BACKGROUND:

Dystrophin Dp71 is one of the isoforms produced by the DMD gene which is mutated in patients with Duchenne muscular dystrophy (DMD). Although Dp71 is expressed ubiquitously, it has not been detected in normal skeletal muscle. This study was performed to assess the expression of Dp71 in human skeletal muscle.

METHODS:

Human skeletal muscle RNA and tissues were obtained commercially. Mouse skeletal muscle was obtained from normal and DMDmdx mice. Dp71 mRNA and protein were determined by reverse-transcription PCR and an automated capillary Western assay system, the Simple Western, respectively. Dp71 was over-expressed or suppressed using a plasmid expressing Dp71 or antisense oligonucleotide, respectively.

RESULTS:

Full-length Dp71 cDNA was PCR amplified as a single product from human skeletal muscle RNA. A ca. 70 kDa protein peak detected by the Simple Western was determined as Dp71 by over-expressing Dp71 in HEK293 cells, or suppressing Dp71 expression with antisense oligonucleotide in rhabdomyosarcoma cells. The Simple Western assay detected Dp71 in the skeletal muscles of both normal and DMD mice. In human skeletal muscle, Dp71 was also detected. The ratio of Dp71 to vinculin of human skeletal muscle samples varied widely, indicating various levels of Dp71 expression.

CONCLUSIONS:

Dp71 protein was detected in human skeletal muscle using a highly sensitive capillary Western blotting system.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Distrofina / Músculo Esquelético Tipo de estudo: Diagnostic_studies Limite: Animals / Humans Idioma: En Revista: Int J Mol Sci Ano de publicação: 2018 Tipo de documento: Article País de afiliação: Japão

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Distrofina / Músculo Esquelético Tipo de estudo: Diagnostic_studies Limite: Animals / Humans Idioma: En Revista: Int J Mol Sci Ano de publicação: 2018 Tipo de documento: Article País de afiliação: Japão