Figure 8.

RagC complexes isolated by immunoprecipitation from Huh7 cells. (A) Huh7 cultured with Si-OH, Si-NH2, or PS-NH2 NPs (all 50 µg/ml) bearing BSA or RNase (both 50 µM) as hard protein corona or bare NPs for 4 h. After 4 h post NP treatment, cells were lysed with lysis buffer from immunoprecipitation kit (Abcam). RagC-mTOR complexes were co-immunoprecipitated from the precleared cell lysates with appropriate Ab as described in the manufacturer’s instruction. The resulting protein complex was eluted from the beads with Laemmli protein sample buffer for SDS-PAGE (Bio-Rad) and resolved on SDS-PAGE with specific antibody against mTOR (Cell Signaling) (full blots are presented in Supporting Information). (B) Scheme of district biochemical signaling activation in cells after stimulation with amino-functionalized non-biodegradable PS and biodegradable silica NPs. ∆mΦ – mitochondrial membrane potential; LMP – lysosomal membrane permeabilization.