Flow cytometric analysis of NFκB in primary CLL cells upon co-culture with M2-10B4 stromal cells

Casein kinase 1 (CK1) has been proved as an efficient approach in the treatment of chronic lymphocytic leukemia (CLL). In this study, we aimed to describe the mechanism-of-action of the CK1 inhibition on the primary CLL cells that were due to their low viability in vitro stimulated by co-culture with M2-10B4 stromal cells that provide activation stimuli mimicking the microenvironment of the lymph nodes (LN). Based on our RNAseq experiment originating from this co-culture model, we were able to see significant downregulation of multiple activation-related pathways in the CLL cells upon CK1 inhibition, including NFκB signalling pathway. Due to this effect on gene expression level, we wanted to inspect whether the NFκB is downregulated also on the protein level. Thus, we utilized the same experimental design with the flow cytometric detection of intracellular NFκB in primary CLL cells that was performed at the endpoint. The provided FCS files originate from 7 CLL patient samples which underwent this experimental set-up with the flow cytometry-oriented endpoint. All samples from the peripheral blood of the CLL patients were taken after signature of informed consent approved by the Ethical Committee of the University Hospital Brno in accordance with the Declaration of Helsinki.