Analysis of HuDe fibroblasts over-expressing TFEB.

<p>A<b>,</b> Immunoblotting of cells transfected with TFEB. Extracts from cells transfected with TFEB or empty vector as control were incubated with an anti-TFEB antibody. As internal control, an anti-βactin antibody was used. B, Reporter activity of HEXA promoter in the presence of TFEB. The wild type and E-box mutated segments −78/−100 were co-transfected with an excess of TFEB expressing plasmid. Vertical bars indicate reporter activity fold induction in the presence of TFEB, with respect to empty vector (set 1). Measures are the mean ± s.d. of three separate experiments, each one in duplicate. C, Hex A and Total Hex enzymatic activity in cell extracts and culture medium of HuDe fibroblasts expressing TFEB. Results are indicated as mU/mg of proteins (specific activity) for cell extracts and in mU/10⁶ cells for cell culture medium. ** P<0.01.