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. 2016 Mar 4;6:22455. doi: 10.1038/srep22455

Figure 1. Effect of mTOR inhibitors on cell viability and mTOR signaling in GBM cells.

Figure 1

(a) LN-18 cells were treated with serial concentrations of rapamycin-RAP, temisirolimus-TEM, torin-TOR and PP-242 for 24 h, 48 h and 72 h and percentage of viable cell count was assessed by MTT assay. Viable count of untreated cells was assumed as 100%. The graphs represent % viable cell count +/− SEM of three similar experiments performed in triplicates. @p-value < 0.05 Untreated vs. inhibitor treatment for 24 h; #p-value < 0.05 Untreated vs. inhibitor treatment for 48 h; *p–value < 0.05 Untreated vs. inhibitor treatment for 72 h. (b) Protein levels of phospho S6K (Thr 389) and total S6K were measured by immunoblotting of total cell lysates of LN-18 cells treated with two concentrations of RAP (5 μM, 10 μM), TEM (2.5 μM, 5 μM), TOR (50 nM, 100 nM) or PP-242 (50 nM, 100 nM) for 24 h. Representative cropped images of two independent experiments. (c) Protein levels of phospho Akt (Ser 473), total Akt, phospho mTOR (Ser2448) and total mTOR were measured by immunoblotting of total cell lysates of LN-18 cells treated with RAP (10 μM), TEM (5 μM), TOR (100 nM) or PP-242 (100 nM) for 24 h. Representative cropped images of three independent experiments. Images show fold change of treated phospho-S6K or -Akt or -mTOR protein expression (normalised with respect to total-S6K or -Akt or -mTOR) relative to untreated samples obtained by densitometry through ImageJ analysis. Full length blots are included in supplementary Fig. S10.