Fig. 7: Effects of ERK1/2i + CDK4/6i on primary MM-cells co-cultured with or without BMSC-CM.
From: Combination therapy targeting Erk1/2 and CDK4/6i in relapsed refractory multiple myeloma
This cohort includes two patients with NRAS mutations, three patients with KRAS mutations, five patients with RAS-WT, and two patients with the t(1;14). A CD138 + PCs were cultured with or without BMSC-CM and treated for 3D and 5D. MM PCs were maintained in 15% fetal calf serum spiked with the serum obtained from autologous peripheral blood of MM patients. Cells were treated with Erk1/2i (Ei), CDK4/6i (Ci), or Erk1/2i + CDK4/6i (Ei + Ci) at the concentrations indicated. The % cell death is indicated with intensity shades of green colors, and the color scale is shown at the right edge of each heatmap. Each concentration of Erk1/2i and/or CDK4/6i was tested in triplicate. The results, plus or minus the standard error of the mean, ranged from +0.01 to +0.7. B Erk1/2i, CDK4/6i, or Erk1/2i + CDK4/6i effects on apoptosis in MM BM PCs were quantified using Annexin V + 7-AAD staining. Unfractionated BM cells were treated at the indicated concentrations, and cell death was monitored 3D after treatment; % of apoptotic events were detected in CD138 gated PCs using FlowJo V10 software. Flow cytometry data that illustrates the gating strategy for the apoptosis assay is shown in supplementary Fig. S5.
