Journal: Oncotarget

Article Title: Lipid rafts promote liver cancer cell proliferation and migration by up-regulation of TLR7 expression

doi: 10.18632/oncotarget.11697

Figure Lengend Snippet: ( A ) Lipid raft and non-raft fractions were isolated using a non-detergent method. Cholesterol concentration in each fraction of Normal and HCC tissues was measured with an Amplex Red cholesterol assay kit. Results were representative of at least three patients in each group. ( B ) Data from panel A were presented as raft (fractions 4–6) and non-raft (fractions 8–10) cholesterol concentration. * P < 0.05, compared with Normal. ( C ) HepG2 cells were treated with 10 mM MβCD or 1μg/ml gardiquimod for 1h and then lipid raft and non-raft fractions were pooled by centrifugation. The highest level of cholesterol was detected in fraction 5 in both HepG2 cells and tissue samples. Only MβCD depleted cholesterol, but gardiquimod resulted in an increase in lipid rafts. Results were representative of three independent experiments.

Article Snippet: Gardiquimod was purchased from Santa Cruz (California, USA). aPPD was provided by Shanghai Innovative Research Centre of Traditional Chinese Medicine (Shanghai, China).

Techniques: Isolation, Concentration Assay, Amplex Red Cholesterol Assay, Centrifugation

Journal: Oncotarget

Article Title: Lipid rafts promote liver cancer cell proliferation and migration by up-regulation of TLR7 expression

doi: 10.18632/oncotarget.11697

Figure Lengend Snippet: ( A ) HepG2 cells were treated with different concentration of gardiquimod for 4 h and 24 h respectively, and the proliferation rate was detected by MTT assay. * P < 0.05, ** P < 0.01, *** P < 0.001 compared to the untreated control ( n = 3). ( B ) HepG2 cells were treated with different concentration of aPPD for 4 h and 24 h respectively, and the cell mortality was detected by MTT assay. * P < 0.05, ** P < 0.01 compared to the untreated control ( n = 3). ( C ) Migration of cells in culture dishes. The edges (dotted lines) of cultured HepG2 cells at 0 and 24 h after scratch were shown as the injury line and the migration line, respectively. The migration area was calculated as regions between the injury and migration lines. ( D ) Quantification of the cell migration rate. The migration rate of HepG2 cells was effectively reduced with the treatment of aPPD compared with gardiquimod. The data were the averages of two repeated experiments with quadruplets for each time, * P < 0.05, ** P < 0.01.

Article Snippet: Gardiquimod was purchased from Santa Cruz (California, USA). aPPD was provided by Shanghai Innovative Research Centre of Traditional Chinese Medicine (Shanghai, China).

Techniques: Concentration Assay, MTT Assay, Control, Migration, Cell Culture

Journal: Oncotarget

Article Title: Lipid rafts promote liver cancer cell proliferation and migration by up-regulation of TLR7 expression

doi: 10.18632/oncotarget.11697

Figure Lengend Snippet: ( A ) HepG2 cells were incubated with 10 mM MβCD or 1 μg/ml gardiquimod for 1 h. Cell lysates were harvested and the proteins were analyzed by immunoblotting with the indicated antibodies. ( B ) TLR7, MyD88, NFκB were quantified and normalized to β-actin. Data were presented as mean ± SEM; n = 3. Statistically significant differences between groups were indicated (* P < 0.05). ( C ) HepG2 cells were treated with 10 mM MβCD or 1 μg/ml gardiquimod for 1h and then lipid rafts (fractions 4–6) were pooled, and immunoprecipitated with anti-flotillin-1 polyclonal antibody, followed by immunoblotting with anti-TLR7 monoclonal antibody. Results were representative of three independent experiments. ( D ) Lipid rafts isolated as described in panel C were immunoprecipitated with anti-TLR7 monoclonal antibody, followed by immunoblotting with anti-flotillin-1 polyclonal antibody. Results were representative of three independent experiments.

Article Snippet: Gardiquimod was purchased from Santa Cruz (California, USA). aPPD was provided by Shanghai Innovative Research Centre of Traditional Chinese Medicine (Shanghai, China).

Techniques: Incubation, Western Blot, Immunoprecipitation, Isolation