Supplemental Figure 1: The table shows the 2-ΔCt values obtained by PCR-array performed with
the cDNA prepared from cocultures of immature MoDC with allogeneic IL-15-activated NK cells
(at 1:1 NK:DC ratio) at 0, 2 and 6 hrs (see Figure 1A). 2-ΔCt values that were upregulated more than
4-fold, or more than 2-fold are highlighted in red or yellow, respectively. PCR array reproducibility
was validated using RT2 Profiler PCR Analysis software (Qiagen). The calculated SD for obtained
Ct-values was ±1.3%. The colour green marks 2-ΔCt values that are downregulated more than 4fold. N.D. = not detected.
Supplemental Figure 2: Lack of expression of activin A by resting and activated NK cells.
Resting NK cells (white bars) and IL-15-treated NK cells (black bars) were cultured in the presence
of IL-2 (200 U/ml), IL-12 (2 ng/ml), IL-15 (40 ng/ml), IL-18 (100 ng/ml), PMA (20 ng/ml) and
ionomycin (5 µM) or on anti-NKp30- or anti-CD16-coated wells for 24 hrs. The dotted line
indicates the ELISA detection limit. Data represent means ±SD of triplicates of one representative
experiment.
Supplemental Figure 3: HPS2 patient-derived NK cells form normal conjugates with DC.
PKH26-labelled MoDC and CFSE-labelled NK cells, isolated from a healthy donor or from a
patient with HPS type 2, were incubated at 37°C for 30 min at a 1:1 NK:DC ratio. Quantification of
NK-DC conjugates (CFSE-PKH26 double-positive) was performed by FACS.
Supplemental Figure 4: Activin A does not affect NK cell cytotoxicity and NK cell receptor
expression. (A) NK cell-mediated DC killing was quantified by enumerating DC after 24 hrs of
coculture with NK cells at the indicated ratios in the presence or absence of 200 ng/ml follistatin.
Data represent mean values ±SD of triplicates of one representative experiment. (B) Autologous
immature DC and activated NK cells were cocultured at a NK:DC ratio of 5:1 in the presence
(tinted histogram) or absence (striped histogram) of 200 ng/ml follistatin for 48 hrs. The expression
of the indicated receptors on NK cells (gated) was analysed by flow cytometry. The dotted
histogram represents the corresponding negative control (only PE-conjugated isotype-specific goatanti-mouse antibody).