Fig. 3
From: Relevance of miR-21 in regulation of tumor suppressor gene PTEN in human cervical cancer cells
Analysis of PTEN protein expression by western blot after miR-21 silencing. Total cellular proteins were obtained from 1 × 105 SiHa cells (HPV16+) per well in a six-well plate containing DMEM at 37 °C with 5 % CO2 after 48 h of transfection with pSilencer1.0-U6 plasmid or pSIMIR21 plasmid (lane 1 cells non transfected NT, lanes 2 and 3 cells transfected with the plasmids, lane 4 (H) corresponds to HaCat cells). The proteins were separated in 12 % SDS-PAGE and were transferred to nitrocellulose membranes which were incubated with each antibody. Panel a corresponds to transfection with pSilencer1.0-U6 plasmid. Panel b corresponds to transfection with pSIMIR21 plasmid and subsequent detection of PTEN protein. Amount similar proteins were analyzed in the immunoblots. The anti-beta-actin antibody was included as control