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Fig. 5 | Journal of Hematology & Oncology

Fig. 5

From: CD73 promotes hepatocellular carcinoma progression and metastasis via activating PI3K/AKT signaling by inducing Rap1-mediated membrane localization of P110β and predicts poor prognosis

Fig. 5

CD73-A2AR axis activates PI3K/AKT signaling by inducing Rap1-mediated membrane localization of P110β. a Rap1 activation status was detected in the indicated HCC cells. b Rap1 expression was silenced via using shRNA in HCCLM3 and SMMC7721 cells and was confirmed by WB assays. c pAKT levels in the indicated HCCLM3 (upper) or SMMC7721 cells (lower) by WB assays. d Cytosolic and plasma membrane fractions were isolated from the indicated HCCLM3 (left) or SMMC7721 cells (right), followed by WB assays with antibody against Rap1. Na+-K+ ATPase was used as an internal control for membrane fractions, and β-actin was used as an internal control for cytosolic fractions. e Co-immunoprecipitation of HA-P110β and Flag-Rap1G12V or Flag-Rap1S12N in SMMC7721 cells. f Effects of the CD73-A2AR axis on promoting Rap1-P110β interaction in co-IP assays. g Cytosolic and plasma membrane fractions were isolated from the indicated HCCLM3 (left) or SMMC7721 cells (right), followed by WB assays with antibody against P110β. pAKT level and PIP3 concentrations were also detected. ELISA experiments for PIP3 determination were conducted in triplicate. h Immunofluorescence staining of Rap1 (green) and P110β (red) with specific antibodies in indicated HCCLM3 (left) or SMMC7721 cells (right). DAPI was used for nuclear staining; scale bar 50 μm. Asterisk indicated P < 0.050 when compared with control group, t tests were used

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