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Since not all ASCL1-high/NEUROD1-low cell lines are sensitive to NB-598, we also included an insensitive cell line (H2081) from this category to more accurately represent the diversity of SCLC cell lines.
Furthermore, growth inhibition against gefitinib insensitive cell lines PC9-GR4 (Del19/tested were tested, compounds 6f and 7e showed about eight and three folds respectively greater potency than gefitinib.
These observations suggest an inherent deficiency in the core components involved in the G2/M progression in the O2 insensitive cell lines.
For example, if SKOV3 and HeyA8 (the O2 insensitive cell lines) proliferate more slowly, O2 dependent proliferation changes may be too trivial to measure.
Similarly, in the O2 insensitive cell lines (HeyA8 and SKOV3) the proportion of mitotic cells remained unaltered regardless of O2 concentrations (Figure 3).
This is expected because, as noted previously (Table 1), the proportion of cells at G2 in the O2 insensitive cell lines were also unaffected by O2 concentration.
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No obvious differences were observed in accumulating lipid droplets between representative SQLE-sensitive and SQLE-insensitive cell lines (Fig. 6b).
Interestingly, SQLE-insensitive cell lines displayed NB-598 sensitivity in the context of serum-free media (Fig. 6e).
To elucidate the differences between SQLE-sensitive and SQLE-insensitive cell lines, we assessed cholesterol biosynthetic pathway status and regulation in SCLC cell lines.
While we observed some differences among cell lines, the baseline level of pathway flux was surprisingly similar between the SQLE-sensitive and SQLE-insensitive cell lines (Fig. 4b).
Interestingly, in the context of abrogated lipid droplets, multiple SQLE-insensitive cell lines displayed sensitivity to NB-598 suggesting that lipid droplets can serve as protective squalene storage depots (Fig. 6d).
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