Fig. 6: Depletion of USP10 increases apoptosis in NSCLC cells upon cisplatin treatment; USP10 confers cisplatin resistance via HDAC6.

a Knockdown of USP10 increases apoptosis upon treatment of cisplatin. H157 cells infected with lentiviruses containing empty vector, shUSP10-1, or shUSP10-2 were treated with cisplatin at the indicated concentrations for 3 days. Cells were lysed, then anti-PARP1, anti-USP10, anti-β-actin and anti-HDAC6 western blotting analyses were performed. b H23 control and USP10 stable knockdown cells were treated with 10 µM cisplatin at the indicated times. Cells were lysed and the anti-PARP1, anti-cleaved caspase 3, and anti-β-actin Western blotting analyses were performed. c Treatment of Dox-induced USP10 knockdown H23 cells with cisplatin increases PARP1 cleavage. H23 cells were pretreated with 1 µg/ml Dox for ~2 days. Cells were then treated with 10 μM cisplatin for the indicated times. Cell lysates were collected for western blotting analyses. d, e Overexpression of USP10 or HDAC6 in USP10-knockdown H1299 cells rescues USP10 knockdown-induced growth reduction and apoptosis upon cisplatin treatment. H1299 control and H1299 USP10 stable knockdown cells, or H1299 USP10 KD cells reintroduced with either USP10 expression plasmid or HDAC6 expression plasmid at each indicated combination, were treated with the indicated concentration of cisplatin for the 3 days. MTT assays were performed in d. Anti-PARP1 and anti-β-actin Western blotting analyses were performed in e. The error bar represents standard deviation. Triple asterisk indicates p < 0.001.