FIG. 6.
ERK5 expression modulates the expression of proangiogenic factors in CCA cells. (A,B) ERK5 knockdown (shERK5) or nontargeting shRNA‐transfected (shNT) cell lysates were used for immunoblotting with antibodies recognizing ERK5, angiopoietin (Ang) 1, or VEGF. Equal protein loading was assessed by vinculin or glyceraldehyde 3‐phosphate dehydrogenase (GAPDH) expression. Densitometries of HuCCT‐1 and CCLP‐1 angiopoietin 1 and VEGF expressions were measured (n = 4) and are shown in the graphs. *P ≤ 0.05 versus shNT. (C) CM was collected from 6 x 105 shNT or shERK5 CCLP‐1 cells, incubated in complete medium for 24 hours, and used for HUVEC tube formation assay. Pictures representative of results obtained with shNT CMs and shERK5 CMs are shown. Tube formation was quantified using ImageJ. *P ≤ 0.05 versus shNT (n = 3).