Table 1.
Experimental designa
| Condition | Mouse strain | Plasmids | Rationale |
|---|---|---|---|
| 1 (nb = 73) | GFP-LC3:GAA−/− | mCherry-LAMP1 | To address lysosomal–autophagosomal interactions; control for conditions 2 & 3 |
| 2 (n = 63) | GFP-LC3:GAA−/− | Flag-TFEB/mCherry-LAMP1 | To investigate the effect of TFEB on lysosomal-autophagosomal clearance |
| 3 (n = 45) | GAA−/− | GFP-TFEB/mCherry-LAMP1 | To study TFEB's effect on lysosomal clearance; control for condition 4 |
| 4 (n = 55) | Atg7:GAA DKO | GFP-TFEB/mCherry-LAMP1 | To study the effect of autophagy on TFEB-induced lysosomal clearance |
| 5 (n = 29) | Atg7:GAA DKO | mCherry-LAMP1 | Control for condition 4 |
| 6 (n = 5) | GFP-LC3:WT | mCherry-LAMP1 | WT control |
| 7 (n = 47) | GAA−/− | GFP-TFEB | To label mitochondria and lysosomes in live and fixed fibres |
a
3–4 animals were used for conditions 1–6; 2 animals were used for condition 7. For each animal ∼1000 fibres were isolated.
b
n = number of fibres analysed by confocal microscopy.