Figure 1

Lentivirus-mediated siRNA inhibited STIM1 expression in U251 cells. (A) Western blot assay: STIM1 protein is expressed in HEK293 cell and human glioblastoma cell lines of different transformation degree, as represented by U373 astrocytoma (WHO Grade III), U87 and U251 glioblastoma multiforme (WHO Grade IV) lines. (B) Transduction efficiency was estimated 72 hrs after transduction at MOI of 50. GFP expression in infected cells was observed under light microscope and fluorescence microscope. Light micrograph (top); Fluorescent micrograph (bottom) (×100). (C) Total RNA was extracted at 72 hrs after transduction and relative STIM1 mRNA expression was determined by quantitative real-time RT-PCR. GAPDH were used to standardize results. Data represent the mean ± S.D. of three independent experiments. **P < 0.01, compared with the si-CTRL group. (D) Total cellular proteins were extracted at 72 hrs after transduction and determined by Western blot analysis using antibodies against STIM1, Orai1, STIM2, with GAPDH as an internal control. Data represent one out of three separate experiments. si-CTRL: cells infected with control-siRNA-expressing lentivirus; si-STIM1: cells infected with si-STIM1.