Effect of growing NCM460 cells in RF deficient and OS media on Sp1 protein and mRNA expression levels and effect of mutating Sp1/GC box on induction in activity of the <i>SLC52A3</i> promoter in RF deficiency.

<p>(A) Western blot analysis was performed using equal amounts (60 μg) of total protein from the cell lysates as described in “Methods”. Blot was probed with the anti-Sp1 polyclonal antibodies. Data represents mean ± SE of four independent sets of experiment are expressed in densitometric values and normalized to the amount of β-actin. Inset is the image of a representative gel. *<i>P</i> < 0.01. (B) The RT-qPCR was performed as described in “Methods” using human Sp1 primers. Data were normalized relative to the human β-actin. Data represents the mean ± SE of five independent sets of experiment *<i>P</i> < 0.01. (C) Activity of the mutated <i>SLC52A3</i> promoter assayed as described in “Methods”. Data are mean ± SE of three independent experiments **<i>P</i> < 0.05.