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. 2014 Oct 29;5(21):10901–10915. doi: 10.18632/oncotarget.2543

Figure 4. Lentivirus-mediated shRNA targeting BRCC3 in U251 and A172 cells.

Figure 4

U251 and A172 cells were infected with lenti-ctrl (mock) or lenti-hBRCC3-shRNA_265 (BRCC3-KD1) and lenti-sh-hBRCC3_731 (BRCC3-KD2). The stable infectants were selected in the presence of puromycin as described in Materials and Methods. (A) The phase-contrast photographs show the morphological change of U251 and A172 cells with BRCC3-KD1 or BRCC3-KD2 toward bipolar or multipolar enlongated cell shapes (arrows). (B) BRCC3 mRNA levels were examined in U251 and A172 cells with BRCC3-KD1 and BRCC3-KD2 using gel-based RT-PCR (upper panel) and QPCR (lower panel). (C) Total proteins were isolated from U251 and A172 cells with BRCC3-KD1 and BRCC-KD2, and then subjected to western blot analysis for the measurement of BRCC3 protein levels. The level of GAPDH was used as a loading control. Data shown are means ± SEM of three independent experiments. **p < 0.01, versus mock (B). Scale bar in A, 200 μm.