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. 2019 Jul 17;10:3137. doi: 10.1038/s41467-019-10948-w

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — CD19 expression profiles on myeloma cells established by dSTORM. a Schematic expression profile of CD19. Density distributions were divided into a CD19-positive subpopulation (CD19-positive cells) and a CD19-negative subpopulation (CD19-negative cells; blue range). The latter group was defined by the density distribution pattern of the isotype control antibody (non-specific binding of the control antibody to the plasma membrane and glass surface). Distributions were fitted to a two-component log-normal function with mean (µ) and standard deviation (σ), to calculate density ranges from small (µ − 2σ) to large (µ + 2σ) values. The CD19-positive population was further divided into a CD19^low (orange range) and a CD19^high subpopulation (red range), based on the cutoff value of 1350 molecules per cell (see text for further details). b, c Expression profiles for CD19 on myeloma cells from patient M012, M016, M019, M022 and a merge c of these four patients generated by dSTORM. Distribution plots show the relative CD19 density obtained after staining with isotype control (left column) and anti-CD19 antibody (right column) on myeloma cells. Densities are provided as logarithmic numbers (natural logarithm, Ln) of molecules per µm². Probability density functions (PDFs) were fitted with a one or two-component log-normal function that was dependent on the fit-accuracy calculated with an Anderson-Darling test (rejected at a p-value < 0.05). For each patient, the percentage of myeloma cells that had been determined to be CD19-positive by flow cytometry is provided for comparison (red segments). Dotted blue line in merge panel: function of the isotype control. Data for all patients are shown in Table 1 and Supplementary Fig. 4. d Mean CD19 molecule density on CD19-positive myeloma cells in molecules per µm² as assessed by dSTORM imaging in n = 10 CD19-positive myeloma patients, and one CD19-negative patient (M014) for comparison. Error bars represent standard deviations of the means from repeated random sampling of the estimated log-normal distribution. e Percentages of CD19-positive and CD19-negative cells, ranging from 10% (M022) to 80% (M019) of CD19-positive cells among patients