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. 2016 Mar 6;7(13):17111–17128. doi: 10.18632/oncotarget.7937

Figure 4. TIA1a promotes cell proliferation in ESCC cells.

Figure 4

(A) KYSE190 and KYSE2270 cells were transiently infected with a mock, TIA1a-, or 1b-expressing retroviruses, and the levels of PARP and exogenous TIA1 (exo) isoforms were measured by western blot analysis using GAPDH as a loading control. (B) KYSE190 cells transiently infected with either a mock-, pTIA1[v1]-FLAG- or pTIA1[v2]-FLAG-expressing retrovirus (1,000 cells/well) were plated in six-well plates and treated with 0.5 mg/mL G418 for two weeks. The colonies in each well were stained with crystal violet, and the colony areas were calculated using ImageJ software. *significantly different from the control value by Student's t test (P < 0.05). (C) For spheroid formation assay, KYSE190 cells transiently transfected with mock or each TIA1 isoform were seeded in ultra-low attachment 96-well round bottom plates and incubated at 37°C for the indicated times (d, days). The areas of spheroids were determined as described in the Materials and Methods section (mean ± SD, n = 8). *significantly different from the control value by Student's t test (P < 0.05). (D) The number of viable cells of each stable transfectant was assessed using a WST assay for the indicated times. The values are expressed as fold changes (mean ± SD, n = 4) compared with the respective control values (0 h). *, significantly different from the control value by Student's t test (P < 0.05). (E) Representative results of FACS analysis using stable transfectants. The raw data were quantified for cell cycle analysis using FACSVerse software.