Figure 2.

HBO1 silencing inhibits OS cell progression in vitro. Stable primary OS cells (pOS-1 and pOS-2) and established OS cell lines (U2OS and MG63) that expressed the HBO1 shRNA (shHBO1-a/shHBO1-b) or the scramble control shRNA (“shC”) were established. After cultured for applied time periods, expression of HBO1 mRNA and listed proteins was shown (A, B, and I); Cell growth (C), viability (D and J), proliferation (E and K), S-phase cell percentage (F), cell migration (G and L) and invasion (H) were tested by the listed assays, with results quantified. For nuclear EdU staining, five random views (n = 5) of total 1, 000 cell nuclei per each condition were used to calculate the average EdU ratio (% vs. DAPI, same for all EdU studies). For “Transwell” and “Matrigel Transwell” assays, five random microscopy views were included to calculate the average number of migrated or invaded cells in each condition (same for all “Transwell” assays). For the in vitro cellular functional studies, the exact same number of viable cells with the applied genetic modifications was initially seeded into each well/dish (“Day-0”/“0h”, same for all figures), and cells were cultured for indicated time periods. “Pare” stands for the parental control OS cells. The data were presented as mean ± standard deviation (SD, n = 5). * P < 0.05 vs. “sh-C” cells. The experiments were repeated five times with similar results obtained. Scale bar = 100 µm (E, G and H).