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. 2021 Nov 17;3(4):zcab044. doi: 10.1093/narcan/zcab044

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© The Author(s) 2021. Published by Oxford University Press on behalf of NAR Cancer.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — Upregulated PARP1 expression in glioma stem cells—impact on PARG inhibitor response. (A) PARP1 mRNA levels from representative glioma stem cells were analyzed by qRT-PCR and normalized to that of normal human astrocytes, (*P < 0.05); insert: Immunoblot analysis of PARP1 among representative GSC cells and normal human astrocyte cells. β-Actin was used as the loading control. (B) PARG mRNA levels from representative glioma stem cells were analyzed by qRT-PCR and normalized to that of normal human astrocytes; insert: immunoblot analysis of PARG among representative GSC cells and normal human astrocyte cells. β-Actin was used as the loading control. (C) Percentage of GSC-83 cells in G1, S or G2/M phase after treatment with vehicle (DMSO), PARGi (2.5 μM), IR (3 Gy) or IR+PARGi (*P < 0.05, **P < 0.01). (D) Relative number of GSC-83 cells (%), normalized to vehicle control, 5-day after treatment: vehicle (DMSO), IR (5 Gy), PARGi (10 μM), IR+PARGi, IR+PARGi+PARPi or IR+PARPi (ABT-888, 10 μM) (**P < 0.01, ****P < 0.0001 related to DMSO). (E) Percentage of non-viable GSC-83 cells on the 5th day after treatment: vehicle (DMSO), IR (5 Gy), PARGi (10 μM), IR+PARGi, IR+PARGi+PARPi or IR+PARPi (ABT-888, 10 μM) (NS = not significant, **P < 0.01, ****P < 0.0001). (F) Relative number of GSC-83 cell following 5-day PARGi treatment at the doses indicated (left) and percentage of non-viable GSC-83 cells (%) after 5-day PARGi treatment (right), at the doses indicated. (G) Relative cell number of GSC-83 cells (left) or GSC-84 cells (right) after a 5-day treatment of DMSO, PARGi (10 μM), PARGi plus PARPi or PARPi (10 μM), normalized to DMSO as 100% (**P < 0.01, ***P < 0.001). (H) After 5-day treatment of GSC-83 cells with DMSO or PARGi (10 μM), cells were washed twice with PBS and then the same number of live cells from each treatment were seeded and cultured in growth medium without any treatment for an additional 5 days. The relative number of cells following the initial PARGi treatment is normalized to the cells seeded from those following the initial DMSO treatment (NS = not significant).