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. 2022 Jul 25;23(15):8171. doi: 10.3390/ijms23158171

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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The effects of metformin, TMZ, and their combination on MGMT proteins in TMZ-resistant LN229 cells. LN229 parent and resistant cells, LN229 (+IRL) and LN229 (+IRL) TMZ-R cells, were treated with the indicated concentrations of metformin, TMZ, and their combination. (A) After 48 h, cell lysates were collected and subjected to Western blotting analysis using antibodies against mtTFA, Twist, and MGMT. ACTN was used as the protein-loading control. The protein bands were quantified through pixel density scanning and evaluated using ImageJ, version 1.44a (http://imagej.nih.gov/ij/) (accessed on 15 July 2022). (B) The ratios of mtTFA/ACTN, TWIST/ACTN, and MGMT/ACTN were plotted in the LN229 (+IRL) and LN229 (+IRL) TMZ-R cells. The results are representative of three independent experiments. # p > 0.05, * p < 0.05, ** p < 0.01, and *** p < 0.001.