Journal: International Journal of Molecular Sciences

Article Title: Ocular Albinism Type 1 Regulates Melanogenesis in Mouse Melanocytes

doi: 10.3390/ijms17101596

Figure Lengend Snippet: The overexpression of OA1 affects the melanin content in an MITF-dependent fashion. Mice OA1 was PCR amplified and then subcloned into the pMSCV PIG vector with Xho I and EcoR I restriction sites. The pMSCV-OA1-GFP plasmid or empty vector was transfected into the mouse melanocytes. The cells were harvested to extract total RNA and total protein. The expression levels of OA1 were qualitatively and quantitatively analyzed by qRT-PCR and Western blot. ( A ) Melanin contents in melanocytes transfected with OA1 , as well as the control; ( B – D ) transfection efficiency of OA1 in mouse melanocytes; ( E – G ) OA1 overexpression influences microphthalmia-associated transcription factor ( MITF ) mRNA and protein levels in mouse melanocytes. Data are shown as the mean ± standard errors ( n = 3 each), * p < 0.05, ** p < 0.01.

Article Snippet: The mice OA1 gene was PCR amplified and then was cloned into the pMSCV PIG vector (Addgene, Cambridge, MA, USA) with Xho I and EcoR I restriction sites.

Techniques: Over Expression, Amplification, Plasmid Preparation, Transfection, Expressing, Quantitative RT-PCR, Western Blot