NPC1 knockout inhibited insulin signalling in CHO cells.

<p>(<b>a</b>) Total cell lysates of K1 CHO cells and 2–2 CHO cells were immunoblotted with anti-NPC1 antibody to determine expression levels of endogenous NPC1 and overexpressed NPC1-YFP. (<b>b</b>) Total cells lysates of K1 CHO cells and 2–2 CHO cells were immunoblotted with anti-Akt and AS160 antibodies to compare expression levels between K1 and 2–2 CHO cells. (<b>c</b>) Total cell lysates from unstimulated and insulin-stimulated cells (100 nM, 20 min) were immunoblotted with phospho-T308 Akt, phospho-S473 Akt, phospho-T642 AS160 and 14-3-3 antibodies to assess insulin signalling.