Journal: Redox Biology
Article Title: S-nitrosylation of HINT1 in macrophages aggravates foam cell formation and atherosclerosis
doi: 10.1016/j.redox.2026.104063
Figure Lengend Snippet: S-nitrosylation of HINT1 aggravates ox-LDL induced foam cell formation through upregulating scavenger receptor A1 and CD36. (A) Representative images and quantification of oil red O staining in HINT1 KO BMDMs transfected with Lenti-WT or Lenti-C84A followed by treatment with or without ox-LDL (100 μg/mL) ( n = 6). Scale bar: 50 μm. (B) Representative images and quantification of lipid droplet staining in HINT1 KO BMDMs transfected with Lenti-WT or Lenti-C84A followed by treatment with or without ox-LDL (100 μg/mL) ( n = 6). Scale bar: 5 μm. (C) Total cholesterol level in HINT1 KO BMDMs transfected with Lenti-WT or Lenti-C84A followed by treatment with or without ox-LDL (100 μg/mL) ( n = 6). (D) Representative images and quantification of Dil-ox-LDL uptake in HINT1 KO BMDMs transfected with Lenti-WT or Lenti-C84A followed by treatment with or without Dil-ox-LDL (40 μg/mL) ( n = 6). Scale bar: 20 μm. (E) Cholesterol efflux to HDL was evaluated in HINT1 KO BMDMs transfected with Lenti-WT or Lenti-C84A and treated with or without ox-LDL (100 μg/mL) ( n = 6). (F) Western blot analysis of SR-A1, CD36 and LOX-1 in HINT1 KO BMDMs transfected with Lenti-WT or Lenti-C84A followed by treatment with or without ox-LDL (100 μg/mL) ( n = 6). (G) Western blot analysis of SR-B1, ABCA1 and ABCG1 in HINT1 KO BMDMs transfected with Lenti-WT or Lenti-C84A followed by treatment with or without ox-LDL (100 μg/mL) ( n = 6). (H) qPCR analysis of SR-A1, CD36 and LOX-1 in HINT1 KO BMDMs transfected with Lenti-WT or Lenti-C84A followed by treatment with or without ox-LDL (100 μg/mL) ( n = 6). (I) qPCR analysis of SR-B1, ABCA1 and ABCG1 in HINT1 KO BMDMs transfected with Lenti-WT or Lenti-C84A followed by treatment with or without ox-LDL (100 μg/mL) ( n = 6). Data are presented as mean ± SEM. One-way ANOVA was used in A–C and E–I. Unpaired t -test was used in D.
Article Snippet: Antibodys against CD36 (18836-1-AP), SR-B1 (21277-1-AP), ABCA1 (26564-1-AP), ABCG1 (13578-1-AP), USF-2 (16614-1-AP), GAPDH (60004-1-lg), β-actin (66009-1-lg), and Tubulin (11224-1-AP) for western blotting, His-tag (66005-1-lg), Flag-tag (20543-1-AP), and HA-tag (51064-2-AP; 66006-2-lg) for western blotting and immunoprecipitation assay were obtained from Proteintech (Chicago, IL, USA); antibody against HINT1 (sc-271790), SR-A1 (sc-166184) for immunoprecipitation assay and immunofluorescence, CD36 (sc-7309) for immunofluorescence, USF2 (sc-293443) for immunoprecipitation assay were obtained from Santa Cruz Biotechnology (CA, USA).
Techniques: Staining, Transfection, Western Blot
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