Research Paper Advance Articles pp 24242—24254

TNFAIP3 ameliorates the degeneration of inflammatory human nucleus pulposus cells by inhibiting mTOR signaling and promoting autophagy

Figure 5. Inhibition of mTOR signaling ameliorates autophagy in the inflammatory human NPCs. (A, B) Torin1 decreases mTOR signaling in inflammatory human NPCs. The phosphorylation level of mTOR at S2448 was measured by Western blot (A). Torin1 treated cells at 20 nM for 24 h. (B) With its quantification normalized to mTOR level. (C, D) Inhibition of mTOR signaling increased autophagy by Western blot for LC3II protein level. (D) With its quantification normalized to GAPDH for TNFAIP3. (E, F) The relative quantification of LC3II and P62 protein. (G) Human NPCs were transfected with adenovirus containing GFP-LC3 and the formation and distribution of GFP-LC3 punctate were observed under confocal microscopy, amplification × 800. Data are represented by the mean ± standard deviation of 3 independent replicates. *** P<0.01, ** P<0.05, * P>0.05 by the Student t-test.