Figure 3.

MIT domain of NRBF2 is required for maturation. (A) Schematic map of full-length and truncated NRBF2-CFP. (B and C) NRBF2 MIT domain is important for regulating SQSTM1 degradation. nrbf2^-/- N2a cells were transiently transfected with CFP, Nrbf2-CFP, dMIT-CFP dCCD-CFP, CFP-MIT or CFP-CCD, the expression of SQSTM1 levels were detected in basal (B) and starvation conditions (EBSS) (C). (D-F) NRBF2 MIT domain is required for regulating autophagosome-lysosome fusion. nrbf2^-/- N2a cells were transiently transfected with CFP, Nrbf2-CFP, dMIT-CFP, dCCD-CFP, CFP-MIT or CFP-CCD in the presence of RFP-GFP-Lc3 transfection. The autophagosome maturation was observed under confocal microscope. Quantification data were presented as the mean ±SEM, n = 20–25 cells from 3 independent experiments. *P < 0.05, **P < 0.01, vs. the relative control. Scale bar: 20 μm