Fig 1: Model foam cells exhibit changes in lipid metabolism-related mRNA expression at different time-points. The mRNA levels of lipid metabolism-related genes, including (A) CD36, (B) PPAR-γ, (C) ABCA1 and (D) ApoE were measured by reverse transcription-quantitative PCR. The mRNA expression levels of CD36, PPAR-γ, ABCA1 and ApoE increased with prolonged ox-LDL induction time compared with those of the control group. At 72 h, the mRNA expression of CD36 in the control group was 1.5 times that of the model group and the mRNA expression of PPAR-γ, ABCA1 and ApoE was highest at 72 h. Therefore, 72 h was chosen as the optimal duration for ox-LDL to induce macrophages to form foam cells. The expression levels in the control group at 0 h, normalized to the housekeeping gene, were set as 1. *P<0.05 compared with the control group at the same time-point. ox-LDL, oxidized low-density lipoprotein; PPAR-γ, peroxisome proliferator activated receptor-γ; ABCA1, ATP-binding cassette transporter A1; ApoE, apolipoprotein E.
Fig 2: ZNF580 inhibits foam cell formation by regulating the expression of lipid-related genes. (A) The expression of ApoE in LV-ZNF580 and Si-ZNF580 foam cells was measured by (a) RT-qPCR and (b) western blot analysis. The expression of (B) CD36, (C) PPAR-γ and (D) ABCA1 in LV-ZNF580 and Si-ZNF580 foam cells was measured by (a) RT-qPCR and (b) western blot analysis. *P<0.05 compared with the control group; #P<0.05 compared with the model group. ZNF580, zinc finger protein 580; LV-ZNF580, lentivirus for ZNF580 overexpression; Si-ZNF580, lentivirus expressing small interfering RNA of ZNF580; PPAR-γ, peroxisome proliferator activated receptor-γ; ABCA1, ATP-binding cassette transporter A1; ApoE, apolipoprotein E; RT-qPCR, reverse transcription-quantitative PCR.
Fig 3: Knockdown of MALAT1 increases ox-LDL uptake, and causes changes in the expression levels of SR-A, SR-B1, ApoE and ABCA1. (A) At 24 h after transfection with NC siRNA or MALAT1 siRNA, macrophages were stimulated with 20 µg/ml Dil-ox-LDL for 4 h and the ox-LDL uptake was observed. Representative fluorescent images. Scale bar, 50 µm. (B) Quantification of mean fluorescence intensity for ox-LDL uptake assay. (C) Relative expression level of MALAT1 in macrophages transfected with NC siRNA or MALAT1 siRNA. (D) Relative mRNA expression levels of SR-A, SR-B1, ApoE and ABCA1 in macrophages transfected with NC siRNA or MALAT1 siRNA for 48 h. (E) Western blotting, and (F) quantification of the relative protein expression levels of SR-A, SR-B1, ApoE and ABCA1 in macrophages transfected with NC siRNA or MALAT1 siRNA for 48 h. Data are presented as the mean ± SD. *P<0.05, **P<0.01, ***P<0.001 vs. NC siRNA group. NC, negative control; siRNA, small interfering RNA; MALAT1, metastasis associated lung adenocarcinoma transcript 1; ox-LDL, oxidized low-density lipoprotein; SR-A, scavenger receptor class A; SR-B1, scavenger receptor class B member 1; ApoE, apolipoprotein E; ABCA1, ATP-binding cassette transporter A1.
Fig 4: TUDCA alleviates lipid storage in macrophage(A) Western blot analysis of the protein expression of ABCA1 and ABCG1 in different TUDCA treatment concentration in THP1 macrophage with oxLDL stimulation (n = 5); t test. Data are represented as mean ± SD.(B) Western blot analysis of the protein expression of ABCA1 and ABCG1 in different TUDCA treatment concentration in RAW264.7 macrophage with oxLDL stimulation (n = 5); t test. Data are represented as mean ± SD.(C) Dil-LDL and oil-red staining of RAW264.7 macrophage. t test. Data are represented as mean ± SD, Scale bars: 10 μm.
Fig 5: APOE isoform-specific effect on gene expression.A, Comparison between APP/E4/Abca1+/- (vehicle plus T0) and APP/E3/Abca1+/- mice (vehicle plus T0). The volcano plot shows differential gene expression between APP/E4/Abca1+/- and APP/E3/Abca1+/- mice. Data were analyzed using EdgeR and the volcano plots are built using p < 0.05 cut-off. Up- and Down-regulated genes are represented in red and blue respectively. On B and C are shown genes that are up- or down-regulated in APP/E4/Abca1+/- mice B, shown is RNA-seq result for genes that are significantly upregulated in APP/E4/Abca1+/- vs APP/E3/Abca1+/- mice. C, shown is RNA-seq result for genes that are significantly down-regulated in APP/E4/Abca1+/- vs APP/E3/Abca1+/- mice. *, p < 0.05, **, p < 0.01, ***, p < 0.001.
Supplier Page from Abcam for Anti-ABCA1 antibody