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. 2021 Oct 12;10(10):68. doi: 10.1038/s41389-021-00357-4

Fig. 4. iP300w suppresses CIC-DUX4 sarcoma cell proliferation in vivo and in vitro.

Fig. 4

A Morphology of NCC-CDS-X1 and Kitra-SRS cell cultures at 48 h of treatment with iP300w (0.3 µM). Scale bar, 50 µm. B ATP assay for cell viability at 48 h of treatment with a serial dilution of iP300w. C NCC-CDS-X1 spheroid morphology after 4 days of treatment with iP300w (0.3 µM). Scale bar, 50 µm. D Immunofluorescence for Ki-67 in NCC-CDS-X1 and Kitra-SRS cells treated for 24 h with iP300w (0.3 µM). Scale bar, 50 µm. E Representative FACS analyses for EdU incorporation in NCC-CDS-X1 cells. Cells were treated with 0.3 µM iP300w for 4, 24, and 48 h. In the last 4 h of treatment proliferating cells were labeled with EdU (10 µM). F Summary of the FACS analyses presented in “E”. Data are presented as mean ± SEM; *p < 0.05, by one-way ANOVA. Results are presented as expression relative to control (n = 3). G Representative FACS analyses for EdU incorporation in NCC-CDS-X1 cells after a pulse of iP300w (0.3 µM). Cells were incubated for different periods with iP300w (0.3 µM) and analyzed by FACS 24 h later. In the last 4 h of the experiment, the cells were incubated with EdU. H Summary of FACS analyses presented in “G”. I Size of the NCC-CDS-X1 xenograft tumors in control and treated mice over 12 days. Mice (n = 8) were treated 1.4 mg/kg iP300w twice daily. J Gross morphology of dissected tumors at the endpoint of the experiment (day 12) presented in “I”. K RT-qPCR for ETV4 and ETV5 in NCC-CDS-X1 and Kitra-SRS cell lines after 4 and 24 h of treatment with 0.3 µM iP300w. Data are presented as mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.001 by one-way ANOVA. Results are presented as expression relative to GAPDH (n = 3).