Fig. 3

Identification of stable knockdown or overexpressing of CP in GB cells. U251-MG and U87-MG GB cell lines were respectively transfected with CP short hairpin (sh)RNA and CP cDNA. To validated clone construction, CP gene expression level was measured by quantitative real-time PCR in stable transfected U251-MG (A) and U87-MG cells (B). GPI-CP expression (membrane-bound CP) is represented in left, while CP all isoforms expression is represented in right (A and B). CP expression is expressed in mRNA expression normalized to their own control ± SEM (n = 3). CP protein level was measured by flow cytometry in stable transfected U251-MG (C) and U87-MG cells (D). GPI-CP expression (membrane-bound CP), obtained without permeabilization, is represented in left graph, while CP all isoforms expression, obtained after permeabilization, is represented in right graph (C and D). CP expression is expressed as RFI ± SEM (n = 5). T-test was performed between normalized Control versus Clones conditions (^*, p-value ≤ 0.05; ^**, p-value ≤ 0.01; ^****, p-value ≤ 0.0001)