Rapamycin administration increases autophagy by decreasing mTOR activity. After transfection with C199-TDP, cells were treated with rapamycin or vehicle as described under “Experimental Procedures.” A, representative Western blots probed with LC3 (to measure autophagy induction) and phospho-p70S6K and total p70S6K (to measure mTOR activity). β-Actin was used as a loading control. B, quantitative assessment of the Western blots shows that, after rapamycin administration, there was a significant increase in the LC3-II/LC3-I ratio, which indicates an increase in autophagy induction. p = 0.0250 as determined by t test analysis. C, quantitative assessment of the Western blots shows that, after rapamycin administration, there was a significant decrease in p70S6K, which indicates a decrease in mTOR activity. p = 0.0343 as determined by t test analysis.