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. 2016 Feb 12;11(2):e0149361. doi: 10.1371/journal.pone.0149361

Fig 3. si-RNA-mediated knockdown of ferritin light chain in glioblastoma multiforme cell lines inhibits cell growth.

Fig 3

(A) Protein blots of samples harvested 72 h after silencing ferritin light chain (FTL) in A172 glioblastoma multiforme (GBM) cells with three respective si-RNAs: si-FTL-1, si-FTL-2, and si-FTL-3. Of these, si-FTL-2 showed the best interference efficiency (si-control group vs. si-FTL-2 group: P < 0.01). “Mock” denotes treatment with transfection reagent only. “si-Control” denotes transfection with scrambled si-RNA as a negative control. GAPDH was used as an internal loading control. The number at the top of each band represents the average densitometric value normalized against that of GAPDH. (B–D) Results of Cell Counting Kit-8 (CCK-8) assays 24, 48, and 72 h after transfection of GBM cells with mock, si-FTL-2, or scrambled control (negative control: NC) si-RNAs. The data points represent the OD ratio of Mock/NC, NC/NC, si-FTL-2/NC at different time points. Data represent the mean ± standard deviation of six replicates per time point. (E) Immunoblot analyses of U251, A172, and U87 GBM cells. Downregulation of FTL resulted in reduced expression of topoisomerase II-alpha (Topo II) and p-Histone H3 (Ser10) (p-His3) in all cell lines (P < 0.01). PCNA expression also decreased in A172 and U87 cells (P < 0.01), but not in U251 cells (p = 0.04). GAPDH was used as an internal loading control. The number at the top of each band represents the average densitometric value from three experiments, normalized against that of GAPDH (FTL, Topo II and PCNA) or His3 (p-His3).