Fig 1: MAP4K4 expression is suppressed by RNAi in BGC-823 cells. (A) The protein expression of MAP4K4 in five GC cell lines (MGC-803, BGC-823, SGC-7901, AGC and MKN-28) was detected by western blotting. GAPDH was used as loading control. The BGC-823 cells demonstrated the highest expression of MAP4K4, and were subsequently selected for further analysis. (B) Western blotting and (C) reverse transcription-quantitative polymerase chain reaction demonstrated the efficiency of MAP4K4 knockdown. **P<0.01, vs. WT group. WT, wild-type; Neg., negative control; Ri-1, Ri-2 and Ri-3, MAP4K4-shRNA-1, -2 and -3; MAP4K4, mitogen-activated protein kinase kinase kinase kinase 4.
Fig 2: Detection of cell proliferation, invasion, and metastasis ability. (A) Transwell assay of normal condition (NC) and silenced MAP4K4 (shRNA-MAP4K4) in MKN-28 and SGC-7901 cell lines; (B) quantification from (A) the two-independent t-tests analyzes the difference of Transwell pictures’ AOD values; (C) wound-healing assay of normal condition (NC) and silenced MAP4K4 (shRNA-MAP4K4) in MKN-28 and SGC-7901 cell lines from 0 h to 24 h. (shRNA is delivered using a vector); (D) relative width of normal condition (NC) and silenced MAP4K4 (shRNA-MAP4K4) in MKN-28 and SGC-7901 cell lines in 24 h compared to 0 h. The two-independent t-tests analyze the difference in relative widths; (E) expression of MAP4K4/Actin determined by Western blot (WB) in MKN-28 and SGC-7901 cell lines before silencing MAP4K4 and after silencing MAP4K4 actin; (F) expressions of E-cadherin (Cat: AF748), N-cadherin (Cat: 13-2100), Vimentin (PRO-309), and control ß-actin proteins determined by WB analysis in MKN-28 and SGC-7901 cell lines before silencing MAP4K4 (NC) and after silencing MAP4K4 (shRNA); (G) relative expression of MAP4K4 with si-ctl or silenced MAP4K4 (shRNA-MAP4K4) in MKN-28 and SGC-7901 cell lines; (H) clone formation assay of MKN-28 and SGC-7901 cells with si-ctl or silenced MAP4K4 (shRNA-MAP4K4); (I) clone formation statistics of MKN-28 and SGC-7901 cells with si-ctl or silenced MAP4K4 (shRNA-MAP4K4). Data of the three independent experiments are represented as mean ± SD. The significance is determined by two-way ANOVA. ** p < 0.01, *** p < 0.001. Correlational analyses are used to obtain the significant differences (Figure 6G,I).
Fig 3: CDC25A affects the protein and mRNA expression levels of the IL-1ß/NIK/NF-?B signaling axis in xenograft tumors. (A) Quantification of the relative expression levels of IL-1ß, NIK and NF-?B mRNA in the KD, NC and control groups. (B) Representative western blot images and (C) quantification of the relative protein expression levels of IL-1ß, NIK and NF-?B in three groups. *P<0.05 vs. NC or control groups. CDC25A, cell division cycle 25A; IL, interleukin; NIK, mitogen-activated protein kinase kinase kinase kinase 4; NF-?B, nuclear factor-?B; KD, knockdown; NC, negative control.
Fig 4: Suppressing the expression of MAP4K4 inhibits the invasiveness of hepatocellular carcinoma cells. An invasion assay of control and MAP4K4 knockdown BGC-823 cells in Matrigel-coated transwell chambers was performed. The cells, which migrated from the upper well of a Transwell chamber into the lower well were (A) stained and images were captured, and (B) subsequently quantified. The data are expressed as the mean ± standard deviation (**P<0.01). WT, wild-type; Neg., negative control; MAP4K4-Ri-3, MAP4K4-shRNA-3 virus-infected cells.
Fig 5: Differentially expressed MAP4K4 and survival differences. (A) Differential expression of MAP4K4 in gastric cancer and normal samples; (B) the expression of MAP4K4 in samples of different pathological stages; (C) the disease-free survival curve of high- and low-MAP4K4-expression groups; (D) the overall survival curve of high- and low-MAP4K4-expression groups; HR, hazard ratio of survival, and p-value was used to check the survival significance between high- and low-MAP4K4-expression groups, whereas the cl values represent the confidence levels; (E) the results of multivariate Cox regression analysis; (F) the results of univariate Cox regression analysis. The lines in plot represent 95% CI of HR.
Supplier Page from Abcam for Anti-MAP4K4/NIK antibody