(A-D) Glioblastoma (GBM) cell lines (LN229 and Gli36) and
patient-derived GBM sphere lines (MGG152) were engineered with a non-targeting
shRNA (shNS) or MSH6-directed shRNA (shMSH6no.1) lentivirus. Immunoblot
confirmed MSH6 knockdown, with Actin as a loading control. Cells were treated
with temozolomide (TMZ), Veliparib/Olaparib or TMZ combination with
Veliparib/Olaparib, and cell viability was evaluated by Cell Titer Glo on day 6.
A: LN229 (TMZ 200 uM, Veliparib 3 uM, Olaparib 1uM) B: Gli36 (TMZ 30 uM,
Veliparib 1 uM, Olaparib 0.5uM) C: MGG152(TMZ 200 uM, Veliparib 3 uM, Olaparib
1uM). * P<0.05, **P<0.001, *** P<0.0001 (student
t-test). (D) Cell viability assay for
Veliparib(left)/Olaparib(right) dose response in LN229shNS and LN229shMSH6no.1.
Cell viability was evaluated by Cell Titer Glo on day 6. (E) Immunoblot showing
loss of MSH6 in MGG123 with MGG4 as a positive control. Cell viability assay to
determine TMZ dose response in MGG123. Cells were treated with specified
concentrations of TMZ, and cell viability was evaluated by Cell Titer Glo on day
6. (F) MGG123 cells were treated with TMZ, Veliparib/Olaparib, or TMZ
combination with Veliparib/Olaparib, and cell viability was evaluated by Cell
Titer Glo on day 6. ** P<0.005 (student t-test). (G)
Cell viability assay for TMZ dose response with/without Veliparib (3.3 uM) in
normal human astrocytes (NHA). Cell viability was evaluated by Cell Titer Glo on
day 6.