Skip to main content
. 2021 Sep 10;297(4):101185. doi: 10.1016/j.jbc.2021.101185

Figure 3.

Figure 3

CRISPR/Cas9-mediated ADAM17 KO blocked VLDLR shedding and its effect on Wnt signaling.A, representative images of Western blotting of sVLDLR in the media, and VLDLR, ADAM10, and ADAM17 in cell lysates of Ad-VLDLRII-infected WT and ADAM17 KO ARPE-19 cells. WT cells and ADAM17 KO cells were transduced with Ad-VLDLRII (MOI = 25) for 24 h. Then, culture media were replaced with serum-free DMEM for another 24 h. After that, the conditioned media and cell lysates were harvested for Western blot analysis. B, quantification of densitometry of sVLDLR in culture media normalized by VLDLR in cell lysates in (A) (n = 3). C, representative images of Western blotting of sVLDLR in culture media, and VLDLR, ADAM10, and ADAM17 in cell lysates in ADAM17 KO cells that were transfected with plasmid overexpressing ADAM17 (pADAM17) or RFP as control. D, quantification of densitometry of sVLDLR in culture media normalized by VLDLR in cell lysates in (C) (n = 3). In A and C, p and m indicated precursor and mature forms of ADAM10 and ADAM17, respectively. Data were presented as mean ± SD. ∗∗∗p < 0.001. ADAM10, a disintegrin and metalloprotease 10; ADAM17, a disintegrin and metalloprotease 17; DMEM, Dulbecco's modified Eagle's medium; MOI, multiplicity of infection; RFP, red fluorescent protein; sVLDLR, soluble ectodomain of VLDLR; VLDLR, very low-density lipoprotein receptor.