Fig. 3: Effects of MT1-MMP on LDLR. | Nature Communications

Fig. 3: Effects of MT1-MMP on LDLR.

From: Membrane type 1 matrix metalloproteinase promotes LDL receptor shedding and accelerates the development of atherosclerosis

Fig. 3

Immunoprecipitation of LDLR (a and d) and MT1-MMP (b and c). Whole-cell lysate from HepG2 (a and b) or Huh7.5 cells (c and d) was subjected to immunoprecipitation using protein G beads and a mouse anti-LDLR (LDLR) or anti-Myc (Myc) antibody (a and d), or protein G beads and a mouse anti-MT1-MMP antibody, a mouse anti-Myc (Myc) antibody or rabbit IgG (RIgG) (b and c). The immunoprecipitated proteins (IP-Beads) and whole-cell lysate (Input) were subjected to immunoblotting. Representative images were shown. Similar results were obtained from three independent experiments. e Confocal microscopy. Huh7.5 cells were fixed, permeabilized, and then incubated with a mouse anti-LDLR monoclonal and a rabbit anti-MT1-MMP monoclonal antibody. MT1-MMP: green; LDLR: red, DAPI: blue. An x-y optical section of the cells illustrates the cellular distribution of proteins (magnification: 100×). f, g MT1-MMP knockdown (n = 3 independent experiments). Primary human hepatocytes were transfected with scrambled (Scram) or MT1-MMP siRNA (MT1). The relative mRNA levels were the ratio of the mRNA levels of the target genes to that of GAPDH at the same condition (f). Same amount of total proteins in whole-cell lysate was subjected to immunoblotting. Cal: calnexin. Relative densitometry was the ratio of the densitometry of LDLR to that of calnexin at the same condition. h Overexpression of MT1-MMP (n = 4 independent experiments). Whole-cell lysate was prepared from primary human hepatocytes infected with empty or the wild-type MT1-MMP (MT1)-adenovirus and then applied to immunoblotting. sLDLR. Medium was collected from primary human hepatocytes treated with siRNA (i) or adenovirus (j) to measure sLDLR with ELISA. Similar results were obtained from at least three independent experiments. Student’s t test (two-sided) was carried out to determine the significant differences between groups (f–j). The significance was defined as p < 0.05. Values of all data were mean ± SD. Source data are provided as a Source Data file.

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