Fig. 1

Expression of NCEH1 in HG-incubated or HFD-induced mouse aortae and HG-exposed ECs. (A) Representative blots and quantitation of NCEH1 protein in NG (mannitol, 25 mM) or HG (D-glucose (25 mM)-incubated mouse aortae. (B) Relative mRNA level of NCEH1 in NG or HG-incubated mouse aortae. (C) NCEH1 activity in NG or HG-incubated mouse aortae. (D) Representative blots and quantitation of NCEH1 protein in NG or HG-incubated ECs. (E) Relative mRNA level of NCEH1 in NG or HG-incubated ECs. (F) NCEH1 activity in NG or HG-incubated ECs. (G) Representative blots and quantitation of NCEH1 protein in normal diet or HFD-incubated mouse aortae. (H) Relative mRNA level of NCEH1 in normal diet or HFD-incubated mouse aortae. (I) NCEH1 activity in normal diet or HFD-incubated mouse aortae. (J) Immunofluorescence double staining showing the expression of NCEH1 in NG or HG-incubated mouse aortae. Scale bar, 200 µm. (K) Immunofluorescence double staining showing the expression of NCEH1 in normal diet or HFD-incubated mouse aortae. Scale bar, 200 µm. n = 4–6. *P < 0.05 versus Control (Con) or normal glucose (NG). The P-value was calculated by unpaired two-tailed Student’s t-test (A-I). For immunoblotting assay, the ratio of the grayscale values of the target protein and β-actin in each group was normalized by the average value of the control group. For RT-PCR assay, the expression levels of the target gene were relative to β-actin and their expression was relatively quantified by the 2^−ΔΔCt method. The ratio of the activity of NCEH1 in each group was normalized to the average value of the control group. NCEH1, neutral cholesterol ester hydrolase 1; NG, normal glucose; HG, high glucose; Con, Control; HFD, high fat diet; DAPI, 4’,6-Diamidine-2’-phenylindole dihydrochloride