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. 2016 May 23;7(26):39980–39995. doi: 10.18632/oncotarget.9556

Figure 1. Progranulin depletion inhibits UMUC-3 urothelial cancer cell motility.

Figure 1

(A) The generation of UMUC-3/shScr (scramble control) and UMUC-3/shPGRN cells has been described in Materials and Methods. The progranulin-specific shRNA was TI350373 (Origine). Progranulin expression in lysates and conditioned media from parental (P), control (Scr) or shRNA progranulin-transfected (shPGRN) UMUC-3 cells was detected by immunoblot with anti-progranulin polyclonal antibodies. (B) Migration of the various UMUC-3 cell lines was performed using transwells as described in detail in Materials and Methods. Data are the average of three independent experiments run in duplicates ± SD. ***P < 0.001. Recombinant human progranulin was supplemented at 80 nM (C) The in vitro “wound healing” motility assay in UMUC-3 cells in either SFM or SFM supplemented with 1% FBS was performed as described in Materials and methods. Cells were analyzed with a cell live microscope using the Metamorph Image Acquisition and Analysis software (Universal Imaging) (×100). 10 fields/plate were examined. Pictures from representative fields are shown.