Journal: Investigative Ophthalmology & Visual Science

Article Title: Dual Inhibition of Histone Deacetylases and the Mechanistic Target of Rapamycin Promotes Apoptosis in Cell Line Models of Uveal Melanoma

doi: 10.1167/iovs.62.12.16

Figure Lengend Snippet: Preliminary screen for drug combinations with romidepsin that promote cell death in uveal melanoma cell lines. ( A ) MP38 cells were initially treated for 6 hours without or with 25 ng/mL romidepsin alone or in combination with pathway inhibitor(s) as indicated. Cells were subsequently incubated in romidepsin-free media in the absence or presence of pathway inhibitors for an additional 42 hours, stained with annexin V and SYTOX green, and analyzed by flow cytometry. Right-most quadrants of each (as indicated by sample portion of first graph boxed in red ) represent annexin-positive cells. ( B ) Heat map containing data for annexin-positive cells for each treatment applied to MP41, Mel202, 92.1, and MP38 cells. The data shown represent the mean of three biological replicates.

Article Snippet: The 92.1 and Mel202 cell lines were purchased from MilliporeSigma (Burlington, MA, USA); MP41 and MP38 cell lines were purchased from American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Incubation, Staining, Flow Cytometry

Journal: Investigative Ophthalmology & Visual Science

Article Title: Dual Inhibition of Histone Deacetylases and the Mechanistic Target of Rapamycin Promotes Apoptosis in Cell Line Models of Uveal Melanoma

doi: 10.1167/iovs.62.12.16

Figure Lengend Snippet: The mTOR inhibitors combined with romidepsin are effective in uveal melanoma cell line models. MP41, Mel202, 92.1, MP38, and ARPE-19 cells were treated for 6 hours with 25 ng/mL romidepsin alone or in combination with 1 µM of rapamycin or NVP-BEZ235 as indicated. Cells were then incubated in romidepsin-free media in the absence or presence of the mTOR inhibitor for an additional 42 hours, stained with annexin V and SYTOX green, and analyzed by flow cytometry. A quantitative analysis of the data is shown. Each bar represents the mean of three biological replicates with the standard deviation indicated by error bars. n = 3, **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05.

Article Snippet: The 92.1 and Mel202 cell lines were purchased from MilliporeSigma (Burlington, MA, USA); MP41 and MP38 cell lines were purchased from American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Incubation, Staining, Flow Cytometry, Standard Deviation