Rg7112

Ruxolitinib, RG-7112, RG-7388, and romidepsin were purchased from Selleckchem. ALRN-6924 was provided by Aileron Therapeutics.

All mouse experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of Rutgers University. WT p53 (p53^+/+, Cata#: 000664), p53-deficient (p53^−/−, Cata#: 002101), CD45.1 (Cata#: 002014) and Apc^Min/+ (Cata#: 002020) mice were obtained from the Jackson Laboratory. IL12-p40-IRES-eYFP mice were a kind gift from Dr. Timothy E. O’Sullivan at UCLA. Mice were housed under a 12-h light/dark cycle with 6 am light on and 6 pm light off. The temperature was maintained between 70° and 74 °F and the humidity was between 30 and 70%. Age- and gender-matched mice at 8–12 weeks old were used for experiments in this study. Animals were randomly assigned to different treatment groups. Sample sizes were chosen based on the power calculation. For TBI treatment, mice were subjected to 12 Gy TBI using a Cs-137 γ-source irradiator at a dose rate of 90 cGy/min. rLIF (Cata#: ESG1107, Millipore), aIL12-p40 antibody (Cata#: 505309, Biolegend), aCD3 antibody (Cata#: BE0001-1, Bio X Cell), IgG (Cata#: BE0091, Bio X Cell), and RG7112 (Cata#: S7030, Selleck Chemicals) were used for mouse treatments. The investigators were blinded to the group allocation during experiments and when assessing outcomes.

HDMECs were incubated with 10 µM of following inhibitors for one hour prior to glucose treatment: Nutlin-3 (cat. no. N6287-5MG; Sigma-Aldrich, Oakville, ON, Canada), or MX69 (cat. no. S8403; Selleckchem, Houston, TX, USA), or RG7112 (cat. no. S7030; Selleckchem, Houston, TX, USA). Cells were subsequently treated with 5- or 30-mM glucose for 6 h and 24 h before cell harvesting.

Isolated cells were expanded to Passage 1 (P1) in monolayer cultures. P1 cells were then seeded at 20,000 or 10,000 cells per well in eight-well chamber slides (Nunc Lab-Tek II Chamber Slide System) and 96- well flat clear bottom black microplates (Corning, NY) respectively. Cells were serum-starved in DMEM with ITS (1X) (Thermo Fisher, Waltham, MA) for 2 hr prior to treatment with 5 μM RG-7112 (Selleck Chemicals, TX), 100 μM o-Vanillin (Sigma-Aldrich, Oakville, ON, Canada) or vehicle (DMSO (0.01%, (Sigma-Aldrich, Oakville, ON, Canada) for 6 hr. Immunocytochemistry was performed as previously described (Cherif et al., 2019 (link)). Apoptosis was detected using a commercial kit (ab176749, Abcam, Cambridge, MA) according to the manufacturer’s instructions. Photomicrographs were acquired with a fluorescent Olympus BX51 microscope equipped with an Olympus DP71 digital camera (Olympus, Tokyo, Japan).