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. 2017 Feb 23;13(2):e1006626. doi: 10.1371/journal.pgen.1006626

Fig 4. TIP60 inhibition reduces stress granule formation in mammalian cells.

Fig 4

A) Decreased acetylation level of histone H4 was detected by Western Blot analysis upon treatment with NU9056. HeLa and MCF7 cells were treated with DMSO or 20 μM NU9056 for 24 hours, and histone extraction and western blot analysis was performed with the indicated antibodies. B) NU9056 does not promote SG formation in HeLa and MCF7 cells. HeLa and MCF7 cells were treated with DMSO or 20 μM NU9056 for 24 hours and immunofluorescences was performed using TIA-1 as marker for SG formation. C) Quantification the average percentage of cells with NU9056-induced SGs. D) Treatment of HeLa and MCF7 cells with NU9056 resulted in a significant decrease of bortezomib-induced SG formation. HeLa and MCF7 cells were treated with DMSO or 20 μM NU9056 for 24 hours, and then 10 μM bortezomib for 4 hours. Immunofluorescence was performed using FMRP, DDX3, and TIA-1 as markers of SG formation. E) Quantification the average percentage of cells with bortezomib-induced SGs. F) Treatment of HeLa and MCF7 cells with NU9056 resulted in a significant decrease of sodium arsenite-induced SG formation. HeLa and MCF7 cells were treated with DMSO or 20 μM NU9056 for 24 hours, and then 100 μM sodium arsenite for 1 hour. Immunofluorescence was performed using FMRP, DDX3, and TIA-1 as markers of SG formation. G) Quantification of the average percentage of cells with sodium-arsenite induced SGs. Results are the average of three independent experiments with at least 500 cells counted per stress granule marker in each independent experiment. Error bar represent the standard error of the mean. * denotes statistical significance at a p-Value < 0.05 determined using an unpaired t-test.