Journal: Life Science Alliance

Article Title: Glioblastoma cellular MAP4K1 facilitates tumor growth and disrupts T effector cell infiltration

doi: 10.26508/lsa.202301966

Figure Lengend Snippet: (A) Representative immunoblot graphs of P-AKT (S473) and AKT levels in MAP4K1-knockdown (KD) U87 and T98G cells and control cells (NC) (n = 3). (B) Representative immunoblot graphs of P-AKT (S473) and AKT levels in MAP4K1 rescue T98G cells and control cells (NC) (n = 3). (C) Representative immunoblot graphs of P-AKT (S473) and AKT levels after treatment with the PI3K inhibitor LY294002 (25 μM) or vehicle DMSO for 48 h in U87 and T98G cells (n = 3). (D, E, F, G) Respective flow cytometry histograms and relative levels of the mean fluorescence intensity of membrane-bound IL-18R and IL-6R after treatment with the PI3K inhibitor LY294002 (25 μM) or vehicle DMSO for 72 h in U87 and T98G cells. (H, I) Respective flow cytometry histograms and relative levels of the mean fluorescence intensity of membrane-bound IL-18R (H) and IL-6R (I) after treatment with LY294002 (25 μM) or vehicle DMSO for 72 h in MAP4K1 rescue T98G cells. (D, E, F, G, H, I) Data are represented as the mean ± SD (n = 3, the data were from an independent experiment, and this experiment was repeated three times), * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 ( t test in (D, E, F, G), one-way ANOVA in (H, I)). Source data are available for this figure.

Article Snippet: The following antibodies and dilutions were used: rabbit polyclonal anti-MAP4K1 (1:1,000, #4472; CST) for immunoblotting, mouse monoclonal anti-P-AKT (S473) (1:2,000, 66444-1-Ig; Proteintech), rabbit polyclonal anti-AKT (1:2,000, #AF6261; Affinity), mouse monoclonal anti-actin (1:10,000, 66009-1; Proteintech), mouse monoclonal anti-GAPDH (1:10,000, 60004-1; Proteintech), goat anti-mouse secondary antibody (1:10,000, SA00001-1; Proteintech), and goat anti-rabbit secondary antibody (1:10,000, SA00001-2; Proteintech).

Techniques: Western Blot, Flow Cytometry, Fluorescence, Membrane