Fig. 1.

Representative dot plots of CLL patient and healthy volunteer (HV) illustrating the flow cytometry analysis method for the identification of B cells with TLR2 expression. a An acquisition gate was established based on FSC and SSC that included mononuclear cells (R1 region). b, d The R1 gated events were analyzed for CD19 PE staining, and the positive cells (CD19⁺) were gated (region R2). The dot plots c, e (CD19 PE vs. TLR2 FITC) were established by the combined gating of events using R1 and R2 regions. The number in the upper right quadrant on the dot plots c, e represents the percentage of CD19⁺/TLR2⁺ cells. Additional analysis for identification of CD19⁺/CD5⁺ cells with TLR2 expression was performed. f, g The R2 gated events (CD19⁺) were analyzed for CD5 PE-Cy5 staining, and the positive cells (CD19⁺/CD5⁺) were gated (region R3). Additionally, CD19⁺/CD5⁻ cells were gated (region R4). The dot plots h, j were established by the combined gating of events using R1, R2 and R3 regions. The dot plots i, k were established by the combined gating of events using R1, R2 and R4 regions. The number in the upper right quadrant on the dot plots h, j represents the percentage of CD19⁺/CD5⁺/TLR2⁺ cells. The dot plots i, k indicate CD19⁺/CD5⁻ cells positive for TLR2