Figure 2.

Identification of stably transfected cells using puromycin. Cells were treated with 600 ng/ml puromycin for 14 days. (A) Observing expression levels of green fluorescence using a fluorescence microscope (magnification, ×100). (B) Flow cytometry was used to evaluate transfection efficiency. Transfection efficiency was 95.05±0.17 and 94.75±0.72% in the H_TLR4-sh2 and NC groups, respectively. The letters “C” and “D” represent the gate of flow cytometry. Measuring expression levels of TLR4 (C) mRNA and (D) protein in transfected cells using fluorescence reverse transcription-quantitative PCR and western blot analysis, respectively. n=3. *P<0.01 vs. U-87MG and U-87MG-NC. TLR4, toll-like receptor 4; sh, short hairpin RNA; NC, negative control; GFP, green fluorescent protein.