Journal: Open Biology

Article Title: The human parasite Leishmania amazonensis downregulates iNOS expression via NF-κB p50/p50 homodimer: role of the PI3K/Akt pathway

doi: 10.1098/rsob.150118

Figure Lengend Snippet: PI3K inhibition reduces p50/p50 occupancy on the iNOS promoter and increases iNOS message levels in infected macrophages. ( a ) RAW 264.7 cells were infected for 5 h with promastigotes of L. amazonensis and treated with ( a ) 10 µM LY294002 (PI3K inhibitor), ( b ) 1 µM wortmannin or ( c ) 5 µM Akti-1/2. The ChIP assay was carried out using anti-p50 antibodies. ( d ) Peritoneal macrophages were infected with L. amazonensis and/or treated with the PI3K inhibitor LY294002 and/or LPS (1 µg ml −1 ). The samples were subjected to qPCR, as previously described. * p < 0.05.

Article Snippet: To inhibit the PI3K/Akt pathway, cells were treated with 10 μM of LY294002 (Sigma-Aldrich) or 1 μM of wortmannin (Sigma-Aldrich) or 5 μM of Akt inhibitor VIII, isozyme-selective, Akti-1/2 (Santa Cruz Biotechnology) during the infection.

Techniques: Inhibition, Infection

Journal: Oncotarget

Article Title: MCL-1-independent mechanisms of synergy between dual PI3K/mTOR and BCL-2 inhibition in diffuse large B cell lymphoma

doi:

Figure Lengend Snippet: Concentrations of PI3K pathway inhibitors used

Article Snippet: We obtained rapamycin, MLN0128, GDC-0941, and NVP-BEZ235 from LC Laboratories (Woburn, MA, USA); ABT-263, ABT-199, MK2206 and GDC-0980 from Active Biochem (Wan Chai, Hong Kong), and AKT inhibitor VIII from Chemdea (Ridgewood, NJ, USA).

Techniques: Concentration Assay

Journal: Frontiers in Oncology

Article Title: Crosstalk between protein kinases AKT and ERK1/2 in human lung tumor-derived cell models

doi: 10.3389/fonc.2022.1045521

Figure Lengend Snippet: Determining the efficacy of targeted drugs by phosphorylation of their targets. (A) Capivasertib, an AKT inhibitor, increases the phosphorylation of AKT itself. Capivasertib efficacy is demonstrated by inhibited phosphorylation of GSK3β, a downstream target of AKT. DM – vehicle control (DMSO), CAP – capivasertib, +cis – cisplatin. (B) AKT inhibitor VIII (AKTi) and PI3K inhibitor idelalisib (IDE) reduce AKT phosphorylation in control and cisplatin-treated cells. (C) MEK/ERK signaling inhibitors suppress ERK1/2 phosphorylation in control and cisplatin-treated cells. SEL – selumetinib, SCH – ERK inhibitor SCH772984, TRA – trametinib. Representative Western blots are shown. Coomassie-stained protein gels are presented as loading controls. 6-hour-long exposures to the drugs were used. The concentration of cisplatin was 90 µM, and that of inhibitors was 10 µM except for trametinib and SCH772984 (1 µM).

Article Snippet: The following inhibitors of ERK and AKT signal pathways were used in this study: PI3K inhibitor idelalisib CAL-101 (10 µM; Cayman Chemical, Ann Arbor, MI, USA); Akt inhibitor VIII (10 µM, Merck) and capivasertib AZD5363 (10 µM, Cayman chemical); MEK1/2 inhibitors selumetinib AZD6244 (10 µM, Selleck Chemicals, Houston, TX, USA), trametinib (1 µM, Cayman chemical); ERK inhibitor SCH772984 (1 µM, Cayman chemical); FAK inhibitor PF573228 (10 µM, Merck.).

Techniques: Western Blot, Staining, Concentration Assay

Journal: Frontiers in Oncology

Article Title: Crosstalk between protein kinases AKT and ERK1/2 in human lung tumor-derived cell models

doi: 10.3389/fonc.2022.1045521

Figure Lengend Snippet: Crosstalk between ERK and AKT (continued). (A, B) AKT inhibitor capivasertib enhances ERK1/2 phosphorylation in control (A) and cisplatin-treated cells (B) . (C) AKT inhibitor VIII enhances ERK1/2 phosphorylation in the cell lines tested, both control and cisplatin-treated cells. Similarly, PI3K inhibitor idelalisib increases ERK phosphorylation in human lung-derived cell lines. Representative Western blots are shown. Coomassie-stained protein gels are presented as loading controls. 6-hour-long exposures to the drugs were used. DM – vehicle control (DMSO), CAP – capivasertib (10 µM), +cis – cisplatin (90 µM), AKTi – AKT inhibitor VIII (10 µM), IDE – idelalisib (10 µM).

Article Snippet: The following inhibitors of ERK and AKT signal pathways were used in this study: PI3K inhibitor idelalisib CAL-101 (10 µM; Cayman Chemical, Ann Arbor, MI, USA); Akt inhibitor VIII (10 µM, Merck) and capivasertib AZD5363 (10 µM, Cayman chemical); MEK1/2 inhibitors selumetinib AZD6244 (10 µM, Selleck Chemicals, Houston, TX, USA), trametinib (1 µM, Cayman chemical); ERK inhibitor SCH772984 (1 µM, Cayman chemical); FAK inhibitor PF573228 (10 µM, Merck.).

Techniques: Derivative Assay, Western Blot, Staining

Journal: Frontiers in Oncology

Article Title: Crosstalk between protein kinases AKT and ERK1/2 in human lung tumor-derived cell models

doi: 10.3389/fonc.2022.1045521

Figure Lengend Snippet: Dependence of the interplay between ERK and AKT signaling pathways on extracellular contacts. (A) Focal adhesion kinase inhibitor PF573228 (+Fi; 10 µM) attenuates the MEK kinase inhibitor selumetinib-induced increase of AKT phosphorylation in control and cisplatin-treated lung cancer-derived cells, except A549 cells. (B) PF573228 prevents the AKT inhibitor capivasertib-induced increase in ERK phosphorylation in control and cisplatin-treated lung cancer-derived cells. (C) Cells grown in suspension (Susp) under agitation, in contrast to adherent cells (Adh), do not show alternative kinase activation after the treatment with inhibitors. Representative Western blots are shown. Coomassie-stained protein gels are presented as loading controls. 6-hour-long exposures to the drugs were used. DM – vehicle control (DMSO), SEL – selumetinib (10 µM), +cis – cisplatin (90 µM), CAP – capivasertib (10 µM), AKTi – AKT inhibitor VIII (10 µM).

Article Snippet: The following inhibitors of ERK and AKT signal pathways were used in this study: PI3K inhibitor idelalisib CAL-101 (10 µM; Cayman Chemical, Ann Arbor, MI, USA); Akt inhibitor VIII (10 µM, Merck) and capivasertib AZD5363 (10 µM, Cayman chemical); MEK1/2 inhibitors selumetinib AZD6244 (10 µM, Selleck Chemicals, Houston, TX, USA), trametinib (1 µM, Cayman chemical); ERK inhibitor SCH772984 (1 µM, Cayman chemical); FAK inhibitor PF573228 (10 µM, Merck.).

Techniques: Derivative Assay, Activation Assay, Western Blot, Staining

Journal: Frontiers in Oncology

Article Title: Crosstalk between protein kinases AKT and ERK1/2 in human lung tumor-derived cell models

doi: 10.3389/fonc.2022.1045521

Figure Lengend Snippet: ERK and AKT crosstalk might be signal strength-dependent. Cells pretreated with a high concentration of cisplatin (240 µM) do not show alternative kinase AKT activation in response to ERK inhibition by selumetinib (A) , nor ERK activation in response to AKT inhibitor VIII, contrary to untreated or low cisplatin concentration (45 µM)-treated cells (B) . Representative Western blots are shown. Coomassie-stained protein gels are presented as loading controls. 6-hour-long exposures to the drugs were used. DM – vehicle control (DMSO), SEL – selumetinib (10 µM), cis - cisplatin, AKTi – AKT inhibitor VIII (10 µM).

Article Snippet: The following inhibitors of ERK and AKT signal pathways were used in this study: PI3K inhibitor idelalisib CAL-101 (10 µM; Cayman Chemical, Ann Arbor, MI, USA); Akt inhibitor VIII (10 µM, Merck) and capivasertib AZD5363 (10 µM, Cayman chemical); MEK1/2 inhibitors selumetinib AZD6244 (10 µM, Selleck Chemicals, Houston, TX, USA), trametinib (1 µM, Cayman chemical); ERK inhibitor SCH772984 (1 µM, Cayman chemical); FAK inhibitor PF573228 (10 µM, Merck.).

Techniques: Concentration Assay, Activation Assay, Inhibition, Western Blot, Staining