Western blots were stripped for hybridization with other primary antibodies in Restore Western Blot Stripping Buffer (Pierce) as specified in the manufacturer's protocol.
10 ml of cell suspension (~1 × 107 parasites) was centrifuged at 2000 rpm in a clinical centrifuge for 10 minutes, washed once for 1 minute with PSG and then the cell pellet lysed in 350 μl RLT buffer containing β-mercaptoethoanol, as specified in the "RNAeasy" RNA preparation protocol (QIAgen).
The remaining reagents (buffer, dNTPs, dithiothreitol, RNase inhibitor, Thermoscript] were added as specified in the Thermoscript protocol.
Mouse BMDC and human monocyte-derived DC (5×104) were treated with 1 µg/mL of the TLR4 agonists MPL, GLA and LPS, or buffer alone (control) for 2 8 h at 37°C as specified in the legends.
Samples were boiled for 5 min in SDS PAGE sample buffer in the presence or absence of reducing agents, as specified in the text and figure legends.
ATP hydrolysis was monitored in real time using the phosphate-sensitive fluorescent sensor MDCC-PBP as described previously (Ferreira et al, 2007) in TSM buffer using 5 nM Chd1 and 100 nM DNA or nucleosomes (as specified in the figures).
The remaining proteins were precipitated using acetone precipitation, as specified in the depletion kit and were re-suspended in buffer containing 7 M urea, 2 M thiourea, 4% CHAPS and 30 mM Tris.
As specified in Section 2.1, the audio frame size is 2,048 samples, the buffer size is 512 samples, and the sampling rate is 44.1 kHz.
Cells were collected, washed with phosphate-buffered saline (PBS), and stained with Annexin V-FiodideoPIdium iodide (PI) (Sigma-Aldrich®) in Annexin-V binding buffer, as specified by the manufacturer.
Timelapse start and duration was as specified in the experiment.
as specified in the Results section.
