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. Author manuscript; available in PMC: 2009 Feb 3.
Published in final edited form as: Nat Chem Biol. 2008 May;4(5):295–305. doi: 10.1038/nchembio.79

Figure 7. Regulation of autophagy by calpain is mTOR-independent.

Figure 7

a, The proportions of EGFP-positive COS-7 cells with aggregates or cell death, after transfection with EGFP-HDQ74 and rheb or pcDNA3.1 (empty vector) (1:3 ratio) for 4 h and treatment with or without 10 μM calpastatin or 50 μM calpeptin for 48 h post-transfection. The control condition for assessing the effect of calpain inhibitors in rheb-transfected cells was taken as 1. Error bars: 95% confidence interval.

b, The proportions of EGFP-positive COS-7 cells with >5 EGFP-LC3 vesicles, transfected with EGFP-LC3 and pcDNA3.1 (empty vector) or rheb (1:3 ratio) for 4 h and treated with or without 10 μM calpastatin or 50 μM calpeptin for 24 h post-transfection. Error bars: 95% confidence interval.

c, The proportions of EGFP-positive COS-7 cells with aggregates, transfected with EGFP-HDQ74 and constitutively active (CA) m-calpain or pcDNA3.1 (empty vector) (1:3 ratio) for 4 h and treated with or without 0.2 μM rapamycin for 48 h post-transfection. Error bars: 95% confidence interval.

d, The proportions of EGFP-positive COS-7 cells with >5 EGFP-LC3 vesicles, transfected with EGFP-LC3 and constitutively active (CA) m-calpain or pcDNA3.1 (empty vector) (1:3 ratio) for 4 h and treated with or without 0.2 μM rapamycin for 24 h post-transfection. Error bars: 95% confidence interval.

e, f, The proportions of EGFP-positive COS-7 cells with aggregates (e) and cell death (f) as in Fig. 1c, treated with or without 0.2 μM rapamycin, 10 μM calpastatin or both for 48 h. Error bars: 95% confidence interval.

g, Clearance of soluble EGFP-HDQ74 in stable PC12 cells as in Fig. 1b, treated with or without 0.2 μM rapamycin, 10 μM calpastatin or both for the 48 h switch-off period. Error bars: Standard error of mean.

h, Clearance of A53T α-synuclein in stable PC12 cells as in Fig. 1a, treated with or without 0.2 μM rapamycin, 10 μM calpastatin or both for the 8 h switch-off period. Error bars: Standard error of mean. ***, p<0.001; **, p<0.01; *, p<0.05; NS, Non-significant.