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. Author manuscript; available in PMC: 2015 Feb 1.
Published in final edited form as: J Immunol. 2014 Jun 27;193(3):1055–1063. doi: 10.4049/jimmunol.1301555

Figure 2.

Figure 2

Changes in plasma cell numbers and functions after addition of bortezomib (2.5 μM), lenalidomide (10 μM) or dexamethasone (10 nM) on days 7 and 11, and assessed 3 days later.

A. Representative plasma cells from cytocentrifuged thymic cells. They were readily identified by their staining for internal IgG (green) and surface CD138 (red; DNA is blue), and typical morphological features, including: relatively large size, abundant cytoplasm with positive staining for immunoglobulins and eccentric nuclei. However, many plasma cells showed dim or negative staining for CD138 despite their intense labeling for internal IgG and typical morphological features (37). They were frequently found in clumps of 2-3 or more cells together, as shown in the lower panel. Scale bars are 10 μm.

B. Normalized plasma cell numbers on day 14. Each point represents a single well (3 per condition) from the indicated patients; horizontal bars are median values. Results are expressed as percentage of the corresponding control cultures of the same patient, from which the absolute plasma cell numbers are given in Table 2.

C. Spontaneous secretion of AChR Abs in vitro, measured by radio-immunoprecipitation and expressed as nanomoles per liter per day (AChR Ab production rate). Each point represents the average of samples from patients MG-1 - MG-3 (4 replicates per patient). Samples from day 14 could not be compared to earlier time points because of inter assay-variation, and are therefore not shown.

D. AChR Ab production rate on day 11. Each point represents a measurement of one well (4 per condition) from the indicated patients; horizontal bars are median values. No AChR Ab production was detected in samples of MG-6.

E. Spontaneous secretion of total IgG in vitro. We measured IgG in supernatants by ELISA, and expressed results as nanograms of IgG per milliliter per day (IgG production).

F. Total IgG production rate on day 14. Each point represents one well (4 per condition) from the indicated patients; horizontal bars are median values. Results are normalized as for Fig 2B, and absolute IgG production rates shown in Table 2.

Arrows indicate the days of drug addition. Error bars correspond to the SEM. One-way (B, D, F) or two-way (C, E) ANOVA and Bonferroni post-hoc testing were used for statistical analyses. Lenalidomide was not tested for MG-8.