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. 2023 Mar 16;24(6):5717. doi: 10.3390/ijms24065717

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© 2023 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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Bcl-2 phosphorylation was regulated by extracellular signal-regulated kinase (ERK). Kasumi-1 and MV4-11 cell lines were treated with 1 µM PD98059, and total protein extracts were analysed by Western blot for the expression of ERK/Perk (A) and pBCL-2(Ser70), pMCL-2 (Ser159/Thr163), and GAPDH expression (B). (C) Kasumi-1 and MV4-11 cell lines were co-treated with 1 µM PD98059 and increasing concentrations (10–1000 nM) of single drugs (ABT-737, Purvalanol-A, AKTi-1/2, and SU9516) for 72 h. Apoptosis was quantified using the Caspase-Glo™ Assay, which quantifies both caspase-3 and -7 activation, indicative of apoptosis. Following treatments, 50 µL of caspase reagent was mixed in equal portion with cell suspension in a 96-well white micotitre plate, incubated at room temperature for 45 min. The luminescence signal was detected using a Synergy HTX Multi-Mode Micro-Plate reader. Data were normalised to vehicle controls and are representative of N = 3 ± SEM.

Bcl-2 phosphorylation was regulated by extracellular signal-regulated kinase (ERK). Kasumi-1 and MV4-11 cell lines were treated with 1 µM PD98059, and total protein extracts were analysed by Western blot for the expression of ERK/Perk (A) and pBCL-2(Ser70), pMCL-2 (Ser159/Thr163), and GAPDH expression (B). (C) Kasumi-1 and MV4-11 cell lines were co-treated with 1 µM PD98059 and increasing concentrations (10–1000 nM) of single drugs (ABT-737, Purvalanol-A, AKTi-1/2, and SU9516) for 72 h. Apoptosis was quantified using the Caspase-Glo™ Assay, which quantifies both caspase-3 and -7 activation, indicative of apoptosis. Following treatments, 50 µL of caspase reagent was mixed in equal portion with cell suspension in a 96-well white micotitre plate, incubated at room temperature for 45 min. The luminescence signal was detected using a Synergy HTX Multi-Mode Micro-Plate reader. Data were normalised to vehicle controls and are representative of N = 3 ± SEM.