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. 2015 Nov 7;108(2):djv326. doi: 10.1093/jnci/djv326

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© The Author 2015. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 4. — Role of ST3Gal1 in glioma cell survival. A) Effect of ST3GAL1 knockdown on common glioma-propagating cell (GPC) markers CD133, CD15, and ALDH; nestin was determined by flow cytometry in NNI-11 (upper panel) and NNI-8 (lower panel) GPCs. NT represents nontargeting vector, while C1 and C2 represent two different shST3GAL1 clones. *P < .05; ***P < .001; vs NT control. For statistical analysis, two-sided Student’s t test was used. (B) Effect of ST3GAL1 knockdown on gliomasphere formation was determined in NNI-11 (upper panel) and NNI-8 (lower panel) GPCs. ***, p<.001; versus NT control at each corresponding day. (C) Effect of ST3GAL1 knockdown on viability was determined in NNI-11 (upper panel) and NNI-8 (lower panel) GPCs. **, P<.01; ***, P<.001; vs NT control at each corresponding day. D, i) The effect of ST3GAL1 knockdown on invasive potential was determined in NNI-11 and NNI-8 GPCs. ***P < .001; vs NT control of corresponding GPCs. D, ii) Cell adhesion proteins p-PAX, PAX, and FAK were evaluated after ST3GAL1 knockdown in NNI-11 (left panel) and NNI-8 (right panel) GPCs. E) Cell cycle profile was (i) evaluated by flow cytometry and (ii) quantified upon ST3GAL1 knockdown in NNI-11 GPCs. *P < .05; **P < .01; vs NT control at each corresponding cell cycle phase. F) Cell cycle profile was (i) evaluated by flow cytometry and (ii) quantified upon ST3GAL1 knockdown in NNI-8 GPCs. *P < .05; **P < .01; vs NT control at each corresponding cell cycle phase. G) Cell cycle proteins were assessed in NNI-11 (left panel) and NNI-8 (right panel) GPCs after ST3GAL1 knockdown and compared with the loading control β-actin. H) Extent of apoptosis, defined by cleaved poly ADP ribose polymerase (PARP) level, and FoxM1 protein levels were determined upon ST3GAL1 knockdown over four to six days in NNI-11 (upper panel) and NNI-8 (lower panel) GPCs. For statistical analysis, unless otherwise specified, one-sided Student’s t test was used, where the assumption was made in advance that the results would only be in one direction (one-sided). This assumption was proven correct by the data. GPCs = glioma-propagating cells.