A good fitness value resulted in increased autolysosomal degradation under ND conditions, as expected.
Baf was included to block lysosomal degradation during both ND-induced autophagy and basal autophagy (FM) (Additional file 1).
Finally, Shafiq et al. demonstrated that PEG-ND is susceptible to light-mediated degradation.
The contact lens device is capable of mediating therapeutic elution from the ND-nanogel via lysozyme-dependent polysaccharide degradation.
This ssDNA generation was Exo1-dependent, as 5' C-strand degradation was undetectable in rif2Δ mre11-nd yku70Δ exo1Δ G1 cells (Figure 4B and 4C), which also failed to accumulate G-strand resection products (Figure 4D and 4E).
Of note, total levels of 4E-BP1 increased under FM in response to BAF, under ND alone, and further with ND/BAF, in accordance with degradation of phosphorylated 4E-BP1 [ 44].
Similarly, the 5' C-strand is not degraded in G1-arrested rif2Δ mre11Δ cells, whereas a residual Exo1-dependent 5' C-strand degradation is detectable in G1-arrested rif2Δ mre11-nd cells.
However, 5' C-strand degradation was undetectable in G2-arrested rif2Δ mre11-nd exo1Δ cells (Figure 3E and 3F), indicating that Exo1 and MRX nuclease activity provide redundant activities to resect the HO-induced telomere in rif2Δ G2 cells.
Similarly, the 5' C-strand was not degraded in G1-arrested rif2Δ mre11Δ cells (Figure 1C and 1D), whereas a residual Exo1-dependent 5' C-strand degradation was detectable in G1-arrested rif2Δ mre11-nd cells (Figure 2C and 2D).
However, compared to FM conditions, under ND conditions with the addition of BAF, a higher rate of autolysosomal degradation was needed in order to obtain a good fit, suggesting that a basal level of lysosomal function occurs under BAF conditions.
