Research Paper Volume 13, Issue 10 pp 14198—14218

Figure 2. Knockdown of LOC146880 inhibits growth and progression of ESCC cells. (A) QRT-PCR analysis shows LOC146880 expression levels in ESCC cells transfected with si-NC (scrambled control siRNA), si-LOC146880#1, si-LOC146880#2, and si-LOC146880#3. (B) Colony formation assay results show viability of Kyse30 and TE-1 cells respectively transfected with si-NC, si-LOC146880#1, or si-LOC146880#2. (C) Immunofluorescence assay results show Ki-67 expression levels in control and LOC146880-silenced Kyse30 and TE-1 cells. (D) 3-dimensional spheroid assay results show the migration ability of control and LOC146880-silenced Kyse30 and TE-1 cells. (E) Wound healing assay results show the migration ability of control and LOC146880-silenced Kyse30 and TE-1 cells. (F) Transwell assay results show the invasiveness of control and LOC146880-silenced Kyse30 and TE-1 cells. (G) Flow cytometry analysis shows apoptotic rates of control and LOC146880-silenced Kyse30 and TE-1 cells. (H) Western blot analysis shows expression levels of E-cadherin (epithelial cell marker) as well as N-cadherin and vimentin (mesenchymal cell markers) in control and LOC146880-silenced Kyse30 and TE-1 cells. (I) Flow cytometry analysis shows cell cycle distribution of control and LOC146880-silenced Kyse30 and TE-1 cells. (J) Western blot analysis shows the levels of pro-apoptotic proteins (cleaved caspase-3 and Bax) and cell cycle proteins (cyclinD1 and CDK4) in control and LOC146880-silenced Kyse30 and TE-1 cells. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001.