Am9855g

To prepare glass milk, 325 mesh silicon dioxide (Spectrum Chemicals, SI108) was combined with an excess volume of 10% HCl (∼3 N HCl) made from combining 37% HCl (Millipore Sigma 320331) and MilliQ water (Millipore) in a fume hood (dry silica powder should not be inhaled). After acid washing for 4 to 8 h at room temperature, silica was pelleted by spinning 2 min at 5,000 × g, and the supernatant was poured off. The pellet was resuspended in four pellet volumes of MilliQ water and then pelleted again. This wash step was repeated for a total of six washes. The pellet was then washed with four pellet volumes of 10 mM Tris HCl, pH = 8 (ThermoFisher Scientific AM9855G) and 1 mM EDTA (ThermoFisher Scientific 15575020), and pelleted. Finally, the pellet was resuspended in one pellet volume of 10 mM Tris HCl and 1 mM EDTA and autoclaved. This autoclave step is likely superfluous, however, as acid washes should render the beads free of contaminants. The resulting 50% glass milk slurry can be stored at room temperature. Before use, care must be taken to vigorously resuspend the particles as they begin to settle quickly.

Lysis buffer: 50 mM Tris-HCl pH 8 (Thermo Fisher AM9855G), 150 mM NaCl (Thermo Fisher AM9760G), 1% Triton X-100 (MilliporeSigma X100-500ML), 0.5% sodium deoxycholate (MilliporeSigma S1827), 0.1% sodium dodecyl sulfate (MilliporeSigma 436143), 1 mM EDTA (Thermo Fisher AM9260G), and 1X cOmplete Protease Inhibitor, EDTA-free (Roche 11836170001) in UltraPure distilled water (Thermo Fisher 10977-015).
After euthanasia as described above, mice were dissected. The relevant tissues were minced to 1 mm pieces, frozen in liquid nitrogen, and stored at –80°C. Chilled lysis buffer was added to the samples in 2 mL microcentrifuge tube. A handheld homogenizer (Fisher Scientific 150, 15-340-167) with a fresh 7 mm plastic probe was used on setting 4 for 10 s to homogenize each sample. Homogenized samples were incubated on ice for 30 min, then centrifuged at 17,000 × g for 10 min at 4°C. The supernatant was transferred to a fresh chilled tube. Normalization, denaturation, SDS-PAGE, western blotting, and quantification were performed as described above for cell culture western blots.

DNA origami structures
were assembled in a single folding reaction carried out in a test
tube (AB0620, ThermoFisher Scientific) with 10 μL of folding
mixture containing 10 nM M13mp18 scaffold DNA (New England Biolabs),
100 nM unmodified oligonucleotides (Integrated DNA technologies),
either 100 nM biotin-modified oligonucleotides (Biomers) for direct
hybridization to the DNA origami tile (H57-dSAv-NL, H57-tSAv-NL) or
500 nM biotinylated oligonucleotides (Biomers) for external hybridization
(H57-dSAv, H57-tSAv) and folding buffer (5 mM Tris (AM9855G, ThermoFisher
Scientific), 50 mM NaCl (AM9759, ThermoFisher Scientific), 1 mM EDTA
(AM9260G, ThermoFisher Scientific), 12.5 mM MgCl2) (AM9530G, ThermoFisher
Scientific)). Oligonucleotide sequences are shown in the Supporting
Information Appendix, Tables S13–S15. At the site chosen for ligand attachment, a staple strand was elongated
at its 3′-end with 21 bases (H57-DNA, H57-PNA, H57-mSAv, H57-dSAv,
H57-tSAv). At sites chosen for cholesterol anchor attachment, staple
strands were elongated at the 5′-end with 25 bases, respectively.
DNA origami were annealed using a thermal protocol (90 °C, 15
min; 90 °C – 4 °C, 1 °C min^–1; 4 °C, 6 h) and purified using 100 kDa Amicon Ultra centrifugal
filters (UFC510096, Merck). DNA origami were stored up to 4 weeks
at −20 °C.