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: Journal of Cancer

Article Title: YTHDF2 inhibit the tumorigenicity of endometrial cancer via downregulating the expression of IRS1 methylated with m 6 A

doi: 10.7150/jca.54527

Figure Lengend Snippet: YTHDF2 regulated the activity of endometrial cell via targeting IRS1. ( A , B ) qRT-PCR and immunoblot analysis of IRS1 in HEC-1-A transfected with negative control (NC), IRS1 siRNA #1 or #2 as indicated for 48 h. ( C , E-H ) qRT-PCR analysis of MMP9 mRNA in HEC-1-A transfected with siRNA ( C,F ), plasmids ( E,G ) or treated with AKT inhibitor ( H ) as indicated, and stimulated with IGF for 6 h. ( D ) Immunoblot analysis of IRS1 in HEC-1-A transfected with the IRS1 plasmids for 48 h. ns: not significant; *p<0.05 (Student's t-test). Data are representative of three independent experiments (mean and s.d. of technical triplicates ( A,C,E-H ).

Article Snippet: AKT kinase inhibitor (HY-10249A) was from MCE (USA).

Techniques: Activity Assay, Quantitative RT-PCR, Western Blot, Transfection, Negative Control

Journal: Blood

Article Title: Lineage-inappropriate PAX5 expression in t(8;21) acute myeloid leukemia requires signaling-mediated abrogation of polycomb repression.

doi: 10.1182/blood-2013-02-482497

Figure Lengend Snippet: Figure 4. Aberrant signaling is required for PAX5 deregulation in t(8;21) AML. (A) Schematic diagram showing chronic AKT, JNK, JAK/STAT, and MAP kinase signaling in t(8;21) AML downstream of an activating mutation (indicated by a star) in a tyrosine kinase (growth factor) receptor or activated rat sarcoma (RAS) signaling. The circles indicate the signaling components targeted by small-molecule inhibitors. (B) Table listing the signaling components targeted, small-molecule inhibitors used, and the effect of the inhibitors on PAX5 expression in Kasumi-1 cells. For detailed data, see supplemental Figure 4A. (C-F) Quantitative reverse transcription PCR experiment measuring expression of PAX5, INK4/ARF, TBP, and RPL13A, respectively, after simultaneous treatment with JNK, MEK, and p38 inhibitors. Each bar graph is representative of 3 independent experiments. The error bars represent the variability in qPCR measurements.

Article Snippet: Cells were treated with 10mM AKT inhibitor (Akt inhibitor IV; Santa Cruz Biotechnology), JNK inhibitor (P600125; Sigma-Aldrich), p38 inhibitor (SB20219; Sigma-Aldrich), MEK inhibitor (PD98059; Sigma-Aldrich), signal transducer and activator of transcription 3 (STAT3) inhibitor (Static; Santa Cruz Biotechnology), or MSK1 inhibitor (H89 dihydrochloride hydrate; Sigma-Aldrich).

Techniques: Mutagenesis, Expressing, Reverse Transcription