Fig 1: FAM49B attenuates the effect of the knockdown of TASP1 on GBC. (A-C) The grow rate and metastasis ability were examined in the Lv-shTASP1 group transfected with/without FAM49B plasmid. (D) The TASP1 and FAM49B expression levels were determined in Lv-shNC and Lv-shTASP1 groups transfected with vector or FAM49B plasmid by western blot. GAPDH was used as the loading control.
Fig 2: TASP1 regulates FAM49B via the PI3K/AKT signaling pathway. (A) The expression levels of TASP1, phosphorylated PI3K (Tyr607)/total PI3K and phosphorylated AKT (Ser473)/total AKT were determined in Lv-shNC and Lv-shTASP1 groups by western blot. (B-D) Myc-TASP1-transfected GBC cells was treated with 20 μM LY294002 for 12 h. The proliferation and migration abilities were reduced. TASP1, phosphorylated PI3K (Tyr607)/total PI3K, phosphorylated AKT (Ser473)/total AKT and FAM49B expression levels were examined by western blot. GAPDH was used as the loading control. (E) The proposed mechanistic scheme of TASP1 regulating FAM49B by activating PI3K/AKT signaling pathway in GBC.
Fig 3: FAM49B is overexpressed and has a positive correlation with TASP1 expression in GBC patients. (A) A portion of cluster analysis of mRNAs that were differential expressed between Lv-shNC- and Lv-shTASP1-transfected GBC-SD cells. Red and blue represent high- and low-expression, respectively. (B) FAM49B expression was downregulated in Lv-shTASP1 group compared with Lv-shNC group. (C) FAM49B expression in GBC tissues and matched non-malignant tissues was evaluated by qRT-PCR. (D) FAM49B expression levels were compared between GBC tissues and their corresponding adjacent tissues. (E-F) The mRNA and protein expression levels of FAM49B were positively correlated with TASP1 expression levels. (G) Kaplan-Meier overall survival curve of patients based on high and low FAM49B expression. (H) The overall survival rates of 72 GBC patients were compared between the high/low TASP1 expression groups and the high/low FAM49B expression groups by Kaplan-Meier analysis.
Fig 4: FAM49B promotes GBC cell proliferation and metastasis. (A) The mRNA expression level of FAM49B was downregulated in GBC-SD and EH-GB-1 cells transfected with Lv-shFAM49B. (B-C) Cell proliferation and colony formation abilities were inhibited in GBC-SD and EH-GB-1 cells transfected with Lv-shFAM49B. (D) Knockdown of FAM49B suppressed the migration and invasion abilities of the GBC-SD and EH-GB-1 cells. (E-F) Statistical significance of colony formation and metastasis assays was analyzed according to the number of colony and invaded cells; **P < 0.01, ***P < 0.001.
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