Fig 1: Screening of the Prestwick Chemical Library corresponding to a collection of 1520 drugs on cell proliferation in the HAP1-KO MBD4 cell line.Scatter plot of the percentage of inhibition of proliferation of HAP1-KO MBD4 cell line treated with a focus on compounds that reduce the proliferation of HAP1-KO MBD4 at over 80%. DMSO was used for the negative control.
Fig 2: Activity of gemcitabine in an MBD4-deficient UM patient-derived xenograft (PDX) model and in the corresponding metastatic patient.A Schema of a co-clinical trial model between a PDX model and the patient from whom was derived the PDX. Created with BioRender.com. B Mice were treated with different drugs and tumor growth was assessed by measuring the relative tumor volume (RTV) for 32 days (n = 4 mice/drug, mean ± SD; n.s non-significant; **P < 0.01; unpaired two-sided Student’s t-test). C Positron emission tomography-computed tomography images before and after 5 months of dacarbazine (upper panel), and before and after 9 months of gemcitabine treatment (lower panel). Arrows show locations of metastatic lesions. D Evolution of the size of metastases during and after dacarbazine or gemcitabine treatment.
Fig 3: Gemcitabine and cytarabine increase cell death of MBD4-deficient cell lines as compared with proficient cell lines.A Analysis of the scatter plot. Annexin V/Propidium Iodide double-negative cells correspond to live cells (blue square), Annexin V positive/Propidium Iodide negative cells correspond to apoptotic cells (green square), and Annexin V/Propidium Iodide double-positive cells correspond to late apoptotic and necrotic cells or undetermined population (orange square). Scatter plots represent a representative experiment among three carried out independently. B Percentage of HAP1 and MEL202, MBD4-proficient (red and green columns) and deficient (blue and violet columns) Annexin V positive cells from three independent assays (mean ± SD; n.s non-significant; *P < 0.05; **P < 0.01; unpaired two-sided Student’s t-test).
Fig 4: MBD4-deficient cell lines are more sensitive to gemcitabine and cytarabine than MBD4-proficient counterparts.A Cell count assay assessing cell viability of HAP1 and MEL02 isogenic cell lines. B IC50 for each culture condition of HAP1 and MEL202 isogenic cell lines was obtained from a dose-response curve (n = 3, mean ± SD). Cell count and IC50 were calculated from three independent assays for each condition. Cytidine addition (2 µM final) or not is indicated. n.s non-significant; **P < 0.01; ***P < 0.001; unpaired two-sided Student’s t-test).
Supplier Page from Abcam for Anti-MBD4/MED1 antibody