Fig. 2: YME1L depletion led to damage of mitochondrial function in primary human NPC cells.

The stable pNPC-1 primary NPC cells with the lentiviral YME1L shRNA (“kdYME1L”), the lentiviral CRISPR-YME1L-KO construct (koYME1L), or the lentiviral scramble control shRNA plus the lentiviral CRISPR-KO control treatment (“ctrl”) were established and expression YME1L (mRNA and protein) was examined (A and B). The exact same number of the above pNPC-1 cells were further cultivated for 36 h, mitochondrial depolarization (via testing JC-1 green monomer intensity, C), ROS levels (by measuring CellROX and DCF-DA fluorescence intensity, D and E) and lipid peroxidation (via testing BODIPY intensity, F) as well as the mitochondrial complex-1 activity (G) and ATP contents (H) were tested. The seahorse assay was performed to measure the oxygen consumption rate (OCR) (I). Primary NPC cells that were derived from other patients, pNPC-2/-3/-4, were stably transduced with the lentiviral YME1L shRNA (“kdYME1L”) or the lentiviral scramble control shRNA (“kdC”), relative expression of YME1L mRNA was shown (J); Cells were further cultivated for 36 h, cellular ATP content (K), mitochondrial depolarization (L), ROS levels (via measuring CellROX fluorescence intensity, M) were tested similarly. The numerical values were mean ± standard deviation (SD, n = 5). “pare” indicates the parental control cells. *P < 0.05 vs. “pare”/“kdC” cells. Experiments in this figure were repeated five times, and each time similar results obtained. Scale bar = 100 μm.