Supplementary Figure Legends

Supplementary Figure 1.

Cellular viability of a) MDA-MB-231, D54MG and T47-D cells after culture for 72h with vehicle or with 10-4 M doxorubicin in serum-free culture medium and b) in MDA-MB-231 cells after culture with vehicleor with 10-6 M taxolin serum-free culture medium, or with vehicle or 10-4 M cis-platinum for 7 days in normal, 10 % FBS-containing culture medium. Data is expressed as % viable cells of total cell counts, as assessed with Trypan blue staining of the cells. Data is expressed as mean ± S.D., n = 3-4, ** p < 0.01, *** p < 0.001 vs. corresponding vehicle treatment.

Supplementary Figure 2.

Cell viability was assessed with MTS-assays in parental MDA-MB-231, or the corresponding stable control or TLR9 siRNA cells after treatment with vehicle or dox DNA/LL-37-complexes (1:10, w/w) for 24 h. Data is expressed as treatment-induced fold-change in viability vs. vehicle (indicated by the red dotted line), mean ± S.E.M., n = 6. No statistically significant differences were detected between the groups.

Supplementary Figure 3.

Stable control or TLR9 siRNA MDA-MB-231 cells were treated for 24 h with vehicle or with the various DNAs, alone or in complex with LL-37 (1:10, w/w). RNA was isolated and a) MMP-2, b) MMP-9, c) MMP-13 or d) TIMP-3 mRNA was quantified with qRT-PCR. Data is expressed as fold-change vs. vehicle (indicated by the dotted line), mean ± S.D., n=4-6, * p<0.05, ** p<0.01, *** p<0.001 vs. corresponding vehicle, ## p < 0.01, ### p < 0.001 vs. corresponding control siRNA cells, ^^ p < 0.01, ^^^ p < 0.001 vs. corresponding treatment without LL-37.

Supplementary Figure 4.

Parental MDA-MB-231 cells were treated in invasion assays with vehicle or with dox DNA: LL-37 complexes (1:10, w/w) alone, or with 50 μM MMP-inhibitor III. Data is expressed as mean ± S.E.M., n=3, * p<0.05, ** p<0.01 vs. vehicle-treatment. The figure is a representative image of 3 separate experiments, and wasconducted also with smaller MMP-inhibitor III concentrations. No inhibition was detected with GM6001 (2 – 25 μM, data not shown) or with specific MMP-9 or MMP-13- inhibitors either, with 5 – 10 μM concentrations (data not shown)

Supplementary Figure 5.

Zymogram results from supernatants of parental MDA-MB-231 cells that were treated with vehicle (veh), the intact (int) or dox DNAs, alone or in complex with LL-37, or with LL-37 alone (the last lane on the right). The gels were developed in the presence of vehicle or MMP-9 inhibitor (5 μM),or with the wide spectrum MMP-inhibitor GM6001 (5 μM). Similar experiments were also performed with MDA-MB-231 cells stably transfected with control or TLR9 siRNAs.

Supplementary Figure 6.Zymogram results from supernatants of a) parental T47-D breast cancer cells as well as the corresponding, stably transfected control and TLR9 siRNA cells and b) parental D54MG glioblastoma cells and the corresponding stable control siRNA and TLR9 siRNA cells. The cells were treated with vehicle (veh), intact (int) or dox DNAs, alone or in complex with LL-37, or with LL-37 alone (the last lane on the right). MW = molecular weight marker.

Supplementary Figure 7.

Cell viability of parental MDA-MB-231 cells was assessed with MTS-assay, after treatment with vehicle or the indicated concentrations of the cathepsin K inhibitorI for 24 h. Data is expressed as mean ± S.E.M., n = 8. No statistically significant differences were detected between the treatmentgroups.

Supplementary Figure 8.

Cell viability of parental MDA-MB-231 cells was assessed with MTS-assays, after treatment with vehicle or the indicated concentrations of doxorubicin for 24 h. Data is expressed as cell viability, % of vehicle (indicated with the red dotted line), mean ± S.E.M.,, n = 6. *** p < 0.001 vs. vehicle, ## p < 0.01 vs. corresponding control siRNA cells.

Supplementary Table 1.

RNA-seq data from vehicle or dox DNA (100 ng/ml) treated MDA-MB-231 cells. The rows high-lighted in red represent genes that were significantly affected by dox DNA-treatment vs. vehicle.

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