Figure 1

RPM contaminate MCSs.

(A) Cells isolated by MCS using CD11c-specific nanoparticles from spleens of Spi-C-deficient and –competent mice, stained for F4/80 and CD11c. (B) Expression of surface markers on RPM identified as shown in (a). Isotype controls given as gray background and autofluorescence (AF) in the 488 nm channel by analyzing unstained cells. (C) RPM contaminations in B and T cells isolated with CD19- or CD3ε-specific mircobeads. (D) Expression of CD19 on B-Cells and RPM (left), as well as expression of CD3ε on CD4 + T cells and RPM (right) in the 650 nm channel. RPM were identified as shown in (a), B cells as B220⁺ CD11c⁻, CD4⁺ T cells as CD4⁺ CD11c⁻. Isotype controls given as gray background. (E) RPM enrichment in splenocytes passed over MCS columns (input) without using paramagnetic nanoparticles, either applying a magnetic field (+Magnet) or not (–Magnet). (F) Ferric iron content of F4/80-negative splenocytes and RPM detected with Prussian blue staining. (G) Colorimetric determination of the iron content of RPM. Depicted is the total iron amount isolated from 1 × 10⁶ RPM according to the manufactures protocol. Results are shown for one representative of two to three individual experiments using 2–4 mice per group. Error bars, s.d. (n = 2–4 mice); *p < 0.05; ***p < 0.001.