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| Figure 2:Down regulation of FTO mRNA expression in SH-SY5Y cells, and FTO’s effect on phospho-Tau levels. (A) SH-SY5Y cells were transfected with siRNA for 24 hours and then incubated for a total of 48 or 72 hours post-transfection resulting in 81.8% and 69.8% knockdown, respectively. Additionally, SH-SY5Y cells were transfected with siRNA and then differentiated for 1 week resulting in 59.3% knockdown. GAPD mRNA was used as a positive internal control. P value: p=0.05. (B) Down regulation of FTO protein expression in SH-SY5Y cells. SH-SY5Y cells transfected with siRNA for 72 hours and immunoblotted post transfection exhibited a 41% decrease in FTO expression. (C) Down regulation of FTO mRNA expression in SH-SY5Y cells. SH-SY5Y cells transfected with siRNA for 24 hours and then incubated for a total of 72 hours post-transfection demonstrated a 69.8% decrease in FTO mRNA expression. (D) FTO knockdown decreases phospho-Tau levels. SHSY5Y cells were either transfected with siRNA and then incubated 48 hours post-transfection, or differentiated for 1 week and subsequently immunoblotted. FTO knockdown decreased the expression of phospho-Tau (Ser396) by 38.1% in naïve and 40.8% in differentiated SH-SY5Y cells. Immunoblots are representative of at least three independent experiments. |
