anti phospho flt3  (Cell Signaling Technology Inc)

Journal: Frontiers in Oncology

Article Title: Targeting TLK2 with antisense oligonucleotides as a new strategy in acute myeloid leukemia

doi: 10.3389/fonc.2026.1659341

Figure Lengend Snippet: TLK2 ASO alone and in combination with gilteritinib decreases viability in AML cell lines. (A) MOLM-14 and MONOMAC-1 cells were treated with TLK2 ASO with a concentration gradient for 7 days, respectively. Cell viability was measured using MTS assay. (B) Within the same experiment (panel A ), MOLM-14 and MONOMAC-1 cells were pre-treated with TLK2 ASO for 4 days and then treated with a FLT3 inhibitor gilteritinib with a concentration gradient for 3 days. Cell viability was measured using MTS assay. IC 50 s were calculated by nonlinear regression in GraphPad Prism 10, fitting normalized cell viability to four-parameter dose-response curves. (C) Immunoblot analysis of signaling in MOLM-14 cells treated with 5 µM NT or TLK ASO, in combination with 20 nM gilteritinib. Statistical significance in panels A and B (*p < 0.05, **p < 0.01, ***p < 0.001) as determined by student t-test.

Article Snippet: Membranes were blocked with 5% BSA in 0.1% Tween20 in 1x PBS (PBS-T) for 1 hr at room temperature, followed by incubation with primary antibodies diluted in 5% BSA in PBST overnight at 4 °C included: TLK2 (#A301-257A, Bethyl Laboratories); Phospho-Stat5 (Tyr694) (#9351, Cell Signaling Technology); Phospho-FLT3 (Tyr589/591) (#3464, Cell Signaling Technology); Actin (#MAB1501, EMD Millipore).

Techniques: Concentration Assay, MTS Assay, Western Blot

Journal: Frontiers in Oncology

Article Title: Targeting TLK2 with antisense oligonucleotides as a new strategy in acute myeloid leukemia

doi: 10.3389/fonc.2026.1659341

Figure Lengend Snippet: TLK2 ASO in combination with gilteritinib decreases leukemic burden in a murine model of AML. (A) A schematic representation of a murine AML bone marrow transplantation model. 1e6 lineage (Lin - )-depleted CD45.2 + LysM-Cre + Flt3 ITD Tet2 +/- bone marrow cells were injected into lethally irradiated CD45.1 + recipients. Mice were treated with NT ASO, TLK2 ASO, NT ASO + gilteritinib, or TLK2 ASO + gilteritinib. (B) TLK2 expression in peripheral blood at week 17. (C) Spleen size measured by ultrasound at 14 weeks post-transplantation. (D) Endpoint spleen weights at 19 weeks post-transplantation. (E) Spleen images from each treatment group. (F) Leukemic burden in the bone marrow at 19 weeks post-transplantation was measured by flow cytometry using CD45.2 + and myeloid marker Ly6GLy6C. (G) Number of Lin - c-Kit + population and Lin - c - Kit + Sca - 1 + population in the bone marrow at week 19. (H) 2, 500 lineage-negative bone marrow cells from LysM-Cre + Flt3 ITD Tet2 +/- mouse were plated in methylcellulose-based media and treated with 10 µM NT ASO or TLK2 ASO, either alone or in combination with 200 nM gilteritinib. Colonies were counted on day 6. Data was presented in duplicates. Statistical significance (*p < 0.05, **p < 0.01, ***p < 0.001) as determined by one-way ANOVA.

Article Snippet: Membranes were blocked with 5% BSA in 0.1% Tween20 in 1x PBS (PBS-T) for 1 hr at room temperature, followed by incubation with primary antibodies diluted in 5% BSA in PBST overnight at 4 °C included: TLK2 (#A301-257A, Bethyl Laboratories); Phospho-Stat5 (Tyr694) (#9351, Cell Signaling Technology); Phospho-FLT3 (Tyr589/591) (#3464, Cell Signaling Technology); Actin (#MAB1501, EMD Millipore).

Techniques: Transplantation Assay, Injection, Irradiation, Expressing, Flow Cytometry, Marker