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. 2021 Mar 11;12(3):259. doi: 10.1038/s41419-021-03542-w

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — A RT-PCR-based fold change determination in expression of various genes known for their role in drug resistance. Imatinib-treated Shh-K562 cells (TShh-K562) and treated K562 (TK562) normalized to their respective untreated controls, at 72 h. Bars represents mean ± SD from three biological repeats. B mRNA-fold change of three anti-apoptotic genes, Bcl2, Mcl-1, Bcl-xL, measured using RT-PCR in Imatinib-treated Shh-K562 (TShh-K562) normalized to Imatinib-treated K562 (TK562). Bars represents mean ± SD from two biological repeats. C Western blot representing expression of Bcl2 and Mcl-1 from untreated and Imatinib-treated cells (K562, Shh-K562, TK562, and TShh-K562). GAPDH is used as loading control. The graph represents densitometric analysis of relative protein levels normalized to untreated K562. Bar represents mean ± SD from three biological repeats. D Time-curve representing viability (MTT assay) of K562 (black dotted line) and Shh-K562 (green dotted line) cell lines after treatment with Imatinib (0.5 µM); K562 (black solid line) and Shh-K562 (green solid line) for Imatinib (0.5 µM) + Cyclopamine (25 nM) treated conditions; gray solid line represents viability of K562 after Cyclopamine treatment (25 nM). Graph represents mean ± SD from three biological repeats. E RT-PCR data determining the level of Bcl2-mRNA in K562 and Shh-K562 after treatment for 72 h with either vehicle control (untreated), Imatinib-alone, or Imatinib + Cyclopamine normalized to their respective K562 controls (black dotted line at 1). Bar represents mean ± SD from two biological repeats. For all assays, P values, *<0.05, **<0.01, and ***<0.001. Statistical analysis done using t-test.