Journal: Cell reports

Article Title: Oxidized mC modulates synthetic lethality to PARP inhibitors for the treatment of leukemia

doi: 10.1016/j.celrep.2023.112027

Figure Lengend Snippet: (A and B) Immunofluorescence of AML cell lines. (A) Representative images of MOLM-13 and THP-1 cells with γH2AX, RAD51, and DAPI after 72-h treatment with vehicle, LAA (250 μM), Olap (1 μM), or LAA + Olap. Scale bar, 25 μm. (B) γH2AX (top) and RAD51 (bottom) focus/cell quantitation. (C) Relative γH2AX (top) and RAD51 (bottom) MFI in MOLM-13 and THP-1 cells treated with LAA and Olap for 72 h. (D–F) Cell cycle dynamics of LAA + Olap-treated cells. (D) Representative density plots of MOLM-13 and THP-1 cells treated for 72 h with vehicle, LAA, Olap (Olap), or LAA + Olap and stained for EdU incorporation and DAPI. (E and F) Relative frequency of MOLM-13 (top) and THP-1 (bottom) cells (E) and γH2AX MFI within each cell cycle gate (F), normalized to vehicle. Mean ± SEM of 3 or more experiments. *p < 0.05, **p < 0.005 (normalized to vehicle), #p < 0.05 (normalized to Olap). (G and H) Cellular proliferation and cycling dynamics. (G) Schematic of BrdU pulse pre-treatment, followed by 72-h treatment with vehicle, LAA, Olap, or LAA + Olap, and EdU chase. (H) Relative frequency of BrdU + (left) and percentage of EdU + BrdU + (right) of THP-1 cells. Representative experiment, mean ± SEM. (J) MOLM-13 and THP-1 cells treated for 72 h with vehicle, LAA, Olap, or LAA + Olap, stained for Annexin V and DAPI, live (double negative), early apoptotic (Annexin V + DAPI − ), or late apoptotic (Annexin V + DAPI + ). Representative experiment, mean + SD. (I) Comet assay performed on MOLM-13 and THP-1 cells treated for 72 h with vehicle, LAA, Olap, or LAA + Olap or for 2 h with 20 μM etoposide. Shown is the tail moment of treated THP-1 (left) and representative fluorescence images of treated MOLM-13 and THP-1 cells (right). Scale bar, 200 μm. (K and L) Quantification of MOLM-13 and THP-1 cells (seeded at 0.2 × 10 6 cells/mL) (K) and relative p21 MFI (L) upon treatment with vehicle, LAA, Olap, or LAA + Olap for 72 h. Mean ± SEM, n = 2 experiments. *p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.00005.

Article Snippet: anti-human RAD51 antibody , B-Bridge International , Cat# 70-001; RRID:AB_2177110.

Techniques: Immunofluorescence, Quantitation Assay, Staining, Single Cell Gel Electrophoresis, Fluorescence

Journal: Cell reports

Article Title: Oxidized mC modulates synthetic lethality to PARP inhibitors for the treatment of leukemia

doi: 10.1016/j.celrep.2023.112027

Figure Lengend Snippet:

Article Snippet: anti-human RAD51 antibody , B-Bridge International , Cat# 70-001; RRID:AB_2177110.

Techniques: Purification, Blocking Assay, Recombinant, Lysis, Staining, DNA Methylation Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Binding Assay, Single Cell Gel Electrophoresis, Software, Protein Array