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Additional file 1: Table S1. Upregulated polypeptides identified by mass spectrometry from A549 proteome in response to PLA-NP with their respective description.

Additional file 1. Transmission electron micrographs, hydrolysis plots, IR spectra and fitting results.

Additional file 8. Proteins bound to AuNP when using 100Â % FCS. The identity of unique proteins identified, via LTQ-Orbitrap mass spectrometry, in AuNP-FCS complexes were identified using http://www.uniprot.org (taxonomy: mammalia); evaluated when incubated in 100Â % FCS; data presented is of unique proteins identified in all 3 biological replicates; this analysis was performed with 3 technical replicates of each biological replicate; for confidence of identification the proteins presented in these graphs are only proteins found in every biological replicate.

Additional file 1. EELS measured at sites of electron dense particulate matter observed in micrographs. Intracellular particles are identified with red lines (A–F), extracellular particles with green lines (E–F), and particles observed during perceived exocytosis with red lines (G–H); Au = gold, Co = control, P = phosphorus.

Additional file 10. General pattern of AuNP-protein interactions â identity of unique proteins. The identity of unique proteins identified, via LTQ-Orbitrap mass spectrometry, in AuNP-FCS complexes were identified using http://www.uniprot.org (taxonomy: mammalia); evaluated when incubated in 10, 55, and 100Â % FCS; data presented is of unique proteins identified in every biological replicate and all 3 serum conditions.

Additional file 5. Immune response of THP-1 cells to NP solvents. Secretion of CCL2 (A-C), TNF-α (D-F), IL-6 (G-I) and IL-1ß (J-L) from PMA-stimulated cells, in response to sodium citrate (2.2 mM stock) or chitosan (0.1 % stock) at dilutions relevant to AuNP exposures; 1 ng/ml LPS was used for co-stimulation; with 1 and 100 ng/ml LPS for controls; results are expressed as cytokine release in pg/ml, R = 3, and each data point represents the mean ± SEM.

Additional file 7. Proteins bound to AuNP when using 55Â % FCS. The identity of unique proteins identified, via LTQ-Orbitrap mass spectrometry, in AuNP-FCS complexes were identified using http://www.uniprot.org (taxonomy: mammalia); evaluated when incubated in 55Â % FCS; data presented is of unique proteins identified in all 3 biological replicates; this analysis was performed with 3 technical replicates of each biological replicate; for confidence of identification the proteins presented in these graphs are only proteins found in every biological replicate.

Additional file 6. Proteins bound to AuNP when using 10Â % FCS. The identity of unique proteins identified, via LTQ-Orbitrap mass spectrometry, in AuNP-FCS complexes were identified using http://www.uniprot.org (taxonomy: mammalia); evaluated when incubated in 10Â % FCS; data presented is of unique proteins identified in all 3 biological replicates; this analysis was performed with 3 technical replicates of each biological replicate; for confidence of identification the proteins presented in these graphs are only proteins found in every biological replicate.

Additional file 4. AuNPs induced pro-inflammatory response of THP-1 cells in different cell activation states. IL-1β release from THP-1 cells in response to treatment with Au_CHIT-H (administered dose of 2.5 µg/ml) for 4 (A and C) or 24 h (B and D), in the presence and absence of 1 ng/ml LPS co-stimulation, and without (A-B) or with (C-D) PMA-priming. Results are expressed as IL-1β release (pg/ml), and each data point represents the mean ± SEM, R = 3. Statistical significance is shown by * = p

Additional file 3. Cytotoxicity assessment of NP solvents in THP-1 cells. Viability (mitochondrial activity) (A-C) and cytotoxicity (LDH release) (D-F) in PMA-stimulated cells, after treatment with sodium citrate (2.2 mM stock) or chitosan (0.1 % stock) at dilutions relevant to AuNP exposures, for 4, 24 and 48 h; 1 ng/ml LPS was used for co-stimulation, Triton X-100 was used as positive control. Results are expressed as fluorescence intensity for viability assays, and as absorbance for LDH release, R = 3, each data point represents the mean ± SEM.

Supplementary Material - Stoehr et al. PFT 2015. (PDF 838Â kb)

Additional file 9. General pattern of AuNP-protein interactions â number of unique proteins. The number of unique proteins identified, via LTQ-Orbitrap mass spectrometry, in AuNP-FCS complexes were identified using http://www.uniprot.orf (taxonomy: mammalia); evaluated when incubated in 10, 55, and 100Â % FCS; data presented is of unique proteins identified in every biological replicate and all 3 serum conditions.

Additional file 2. Au_CHIT-L induced cytotoxicity in THP-1 cells. Viability (mitochondrial activity) (A) and cytotoxicity (LDH release) (B) in PMA-stimulated cells, after treatment with Au_CHIT-L, for 24 and 48 h; 1 ng/ml LPS was used for co-stimulation, Triton X-100 was used as positive control. Results are expressed as, for viability, % viability compared to 100 % control cells, and as the ratio change compared to controls for LDH release, and each data point represents the mean ± SEM, R = 4; statistical significance (determined by ANOVA with Tukey posthoc) is shown by ^ = p

Additional file 2: Table S2. Downregulated polypeptides identified by mass spectrometry from A549 proteome in response to PLA-NP with their respective description.

Determination of conjugated allergen after the incubation in human plasma performed by Coomassie-stained SDS-PAGE (A-C) and Western blots (D-F). (TIF 526Â kb)

Additional file 2: Table S2. Downregulated polypeptides identified by mass spectrometry from A549 proteome in response to PLA-NP with their respective description.

Determination of conjugated allergen after the incubation in human plasma performed by Coomassie-stained SDS-PAGE (A-C) and Western blots (D-F). (TIF 526Â kb)