In the experiment, different levels of APS concentration were added to the solution (0.4 % polymer + 0.4 % cross-linker).
PAC (5, 10 ng/ml) and PAC-csMSN (equivalent to 5 and 10 ng/ml of PAC concentration) were added and incubated for 48 h.
After 20 to 24 h, the RNase A@C-dots and C-dots of certain concentration were added into the E-plates to mix with cells.
The lysates were centrifuged at 14,000×g at 4 °C for 15 min, and sodium dodecyl sulfate (SDS) and sodium deoxycholic acid (0.2% final concentration) were added.
The PS-QD micelles PS (0), (40), (50), (60), and (100) at 10-nM concentration were added to the cells and incubated for 4h at 37°C.
Impurity atom(s) of various characteristics and concentration were added to the near vicinity of the grain boundary to determine the structural and energetic affect.
The culture medium was removed and PS-QD micelles PS (0), (40), (50), (60), and (100) at 10-nM concentration were added and incubated for 4h at 37°C.
Test compounds or vehicle (0.375% ethanol [v·v− 1], final concentration) were added to the cells seeded in 96-well plates and samples were incubated for 30 min at 37 °C; positive control for cell lysis: 1% triton or 16.6% ethanol (v·v− 1).
The 4 ml solution of 1% sodium citrate, Tris, and glucose with the same mass ratio of the concentration were added, respectively, to three parts of 97 ml chloroauric acid solution of concentration 0.01% (in which the volume with trisodium citrate gold solution was 100 ml, since the reaction does not require adding the alkaline solution).
To create β2GPI/anti-β2GPI complexes, anti-β2GPI antibodies at constant concentration were added to β2GPI before the samples were applied to cardiolipin.
One hundred microlitres of each concentration were added to LPS coated and uncoated wells (for subtraction of non-specific results) of nitrocellulose microtiter plates (Millipore, USA) and incubated overnight at 37°C in a 5% CO2 incubator.
