Journal: Journal of Lipid Research

Article Title: Apolipoprotein A1 deficiency increases macrophage apoptosis and necrotic core development in atherosclerotic plaques in a Bim-dependent manner

doi: 10.1016/j.jlr.2025.100782

Figure Lengend Snippet: Effect of genetic deletion of ApoA1 (apolipoprotein A1) in low density lipoprotein receptor ( Ldlr ) KO mice on atherosclerotic plaque and necrotic core sizes and levels of plaque apoptosis. Male 10-week-old ApoA1 WT/WT Ldlr KO/KO ( Ldlr KO/KO ) and ApoA1 KO/KO Ldlr KO/KO mice (n = 22, 20) were fed a high-fat diet for 10 weeks. A: Representative images of oil red O (ORO) stained (top row), and hematoxylin and eosin (H&E) stained (bottom row) atherosclerotic plaques from aortic sinus cross sections of Ldlr KO/KO and ApoA1 KO/KO Ldlr KO/KO mice. Dotted black line represents a necrotic core region. B: Quantification of peak plaque area taken at the apex of aortic sinus plaque profiles. Black data points represent the subset of samples used for subsequent immunofluorescent analysis. C: Quantification of necrotic core area relative to peak plaque area. D: Representative images of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) labeled (top row), cleaved-caspase-3 (CC3) stained (middle row), and Mac-3 and Bim coimmunostained atherosclerotic plaques of Ldlr KO/KO and ApoA1 KO/KO Ldlr KO/KO mice. Yellow dashed line outlines the plaque area. “L” represents the lumen of the aortic valve leaflet. E: Quantification of TUNEL-positive nuclei within the atherosclerotic plaque (n = 13, 13). F: Quantification of CC3-positive area relative to the lesional area (n = 13, 13). G: Quantification of Mac-3-positive area relative to the lesional area (n = 13, 13). H: Quantification of Bim-positive area relative to Mac-3 lesional area (n = 13, 13). For B–G: statistical analysis was conducted using an unpaired t test. For H: statistical analysis was conducted using Mann-Whitney test. Statistical significance is considered when P < 0.05. Data represent mean ± SEM. Bim, B-cell lymphoma 2 [Bcl-2] interacting mediator of cell death; CC3, cleaved caspase 3; LDLR, low-density lipoprotein receptor.

Article Snippet: Cells were incubated for 24 h with the absence or presence of 50 μg/ml of human HDL (J64903, Alfa Aesar, Haverhill, MA or 12-16-080412, Athens Research and Technology, Athens, GA) or 50 μg/ml of human ApoA1 (16-6-120101, Athens Research and Technology) along with differing apoptotic inducers including oxLDL (100 μg/ml, J65591, Alfa Aesar), 7-ketocholesterol (10 μg/ml; Cayman Chemical, Ann Arbor, MI), or tunicamycin (TN; 10 μg/ml; Cayman Chemical).

Techniques: Staining, TUNEL Assay, Labeling, MANN-WHITNEY

Journal: Journal of Lipid Research

Article Title: Apolipoprotein A1 deficiency increases macrophage apoptosis and necrotic core development in atherosclerotic plaques in a Bim-dependent manner

doi: 10.1016/j.jlr.2025.100782

Figure Lengend Snippet: Bim deletion in bone marrow of atherosclerotic ApoA1 KO/KO Ldlr KO/KO mice reduces atherosclerotic plaque, necrotic core sizes, and plaque apoptosis. Thioglycolate-elicited mouse peritoneal macrophages (MPM) from wild-type (WT) C57/BL6 and Bim KO/KO mice were harvested and cultured in media containing lipoprotein-deficient serum and treated with or without 50 μg/ml human high-density lipoprotein (HDL) and with or without 10 μg/ml tunicamycin (TN). A: Representative images of CC3 stained peritoneal macrophages from WT mice (top row) and Bim KO/KO mice (bottom row). B, C: Quantification of CC3 stained MPMs from WT mice and Bim KO/KO mice, respectively (n = 4, 4). Isolated bone marrow from WT mice and Bim KO/KO mice were transplanted into lethally irradiated male 10-week-old Ldlr KO/KO . Following a 4-week recovery period post transplantation, transplanted mice were fed a high-fat diet (HFD) for 10 weeks. D: Representative images of hematoxylin and eosin (H&E) stained atherosclerotic plaques from transplanted Ldlr KO/KO mice. Dotted black line represents the necrotic core region. Analysis of E: peak plaque area, and F: relative necrotic core from transplanted Ldlr KO/KO mice (n = 11, 7). Black data points represent the subset of samples used for subsequent immunofluorescent analysis. G: Representative images of TUNEL (left column) and CC3 stained (right column) atherosclerotic plaques of transplanted Ldlr KO/KO mice. Yellow dashed line outlines the plaque area. “L” represents the lumen of the aortic valve leaflet. H: Quantification of TUNEL-positive nuclei within the atherosclerotic plaque (n = 6, 6). I: Quantification of CC3-positive area relative to the lesional area (n = 6, 6). Male 10-week-old ApoA1 KO/KO Ldlr KO/KO mice were transplanted with BM from WT or Bim KO/KO donors and fed a HFD for 10 weeks in a similar fashion as above Ldlr KO/KO mice. J: Representative images of H&E-stained atherosclerotic plaques from transplanted ApoA1 KO/KO Ldlr KO/KO mice. Analysis of K: peak plaque area, and L: relative necrotic core from transplanted ApoA1 KO/KO Ldlr KO/KO mice (n = 15, 13). M: Representative images of TUNEL labeled (left) and CC3 stained (right) atherosclerotic plaques of transplanted ApoA1 KO/KO Ldlr KO/KO mice. N: Quantification of TUNEL-positive nuclei within the atherosclerotic plaque (n = 13, 10). O: Quantification of CC3-positive area relative to the lesional area (n = 13, 10). For B, C: statistical analysis was conducted using one-way ANOVA with Tukey's post hoc multiple comparisons test. For E, L, O: statistical analysis was conducted using Mann Whitney test. For F–K and N: statistical analysis was conducted using an unpaired t test. Statistical significance is considered when P < 0.05. Data represent mean ± SEM. ApoA1, apolipoprotein A1; Bim, B-cell lymphoma 2 [Bcl-2] interacting mediator of cell death; CC3, cleaved caspase 3; LDLR, low-density lipoprotein receptor; MPM, mouse peritoneal macrophage.

Article Snippet: Cells were incubated for 24 h with the absence or presence of 50 μg/ml of human HDL (J64903, Alfa Aesar, Haverhill, MA or 12-16-080412, Athens Research and Technology, Athens, GA) or 50 μg/ml of human ApoA1 (16-6-120101, Athens Research and Technology) along with differing apoptotic inducers including oxLDL (100 μg/ml, J65591, Alfa Aesar), 7-ketocholesterol (10 μg/ml; Cayman Chemical, Ann Arbor, MI), or tunicamycin (TN; 10 μg/ml; Cayman Chemical).

Techniques: Cell Culture, Staining, Isolation, Irradiation, Transplantation Assay, TUNEL Assay, Labeling, MANN-WHITNEY

Journal: Journal of Lipid Research

Article Title: Apolipoprotein A1 deficiency increases macrophage apoptosis and necrotic core development in atherosclerotic plaques in a Bim-dependent manner

doi: 10.1016/j.jlr.2025.100782

Figure Lengend Snippet: Bim deletion in bone marrow-derived cells of atherosclerotic mice reduces plasma cholesterol and increases circulating leukocytes. Peripheral blood was collected by cardiac puncture from 10-week HFD-fed WT and Bim KO/KO BMT mice fasted overnight. Absolute concentrations of (A) leukocytes; (B) B cells; (C) T cells; and (D) monocytes in peripheral blood samples of LDLR KO/KO BMT mice (n = 9, 5) measured by flow cytometry. Absolute concentrations of (M) leukocytes; (N) B cells; (O) T cells; and (P) monocytes in peripheral blood of ApoA1 KO/KO Ldlr KO/KO BMT mice (n = 8, 7). E and Q, Ratio of Ly6C High (Hi) monocytes to Ly6C Low (Lo) monocytes in Ldlr KO/KO BMT and ApoA1 KO/KO Ldlr KO/KO BMT mice, respectively. F and R, Plasma IL-6 concentration in Ldlr KO/KO BMT (n = 5, 3) and ApoA1 KO/KO Ldlr KO/KO BMT mice (n = 13, 10), respectively. Lipids were analyzed by performing colorimetric enzymatic assays on plasma samples isolated from collected blood. Quantification of G, total; H, free; I, esterified; J: HDL, K: non-HDL cholesterol; and L, triglycerides from plasma of Ldlr KO/KO BMT mice (n = 11, 7). Quantification of S, total; T, free; U, esterified; V, HDL, W, non-HDL cholesterol; and X, triglycerides from plasma of ApoA1 KO/KO Ldlr KO/KO BMT mice (n = 16, 14). For A, C, D, F–I, K, M–P, S–U, and W, statistical analysis was conducted using unpaired t test. For B, E, J, L, Q, R, V, and X: statistical analysis was conducted using Mann Whitney test. Statistical significance is considered when P < 0.05. Data represent mean ± SEM. Bim, B-cell lymphoma 2 [Bcl-2] interacting mediator of cell death; BMT, bone marrow transplantation; HFD, high-fat diet; IL-6, interleukin-6; LDLR, low-density lipoprotein receptor.

Article Snippet: Cells were incubated for 24 h with the absence or presence of 50 μg/ml of human HDL (J64903, Alfa Aesar, Haverhill, MA or 12-16-080412, Athens Research and Technology, Athens, GA) or 50 μg/ml of human ApoA1 (16-6-120101, Athens Research and Technology) along with differing apoptotic inducers including oxLDL (100 μg/ml, J65591, Alfa Aesar), 7-ketocholesterol (10 μg/ml; Cayman Chemical, Ann Arbor, MI), or tunicamycin (TN; 10 μg/ml; Cayman Chemical).

Techniques: Derivative Assay, Clinical Proteomics, Flow Cytometry, Concentration Assay, Isolation, MANN-WHITNEY, Transplantation Assay

Journal: Journal of Lipid Research

Article Title: Apolipoprotein A1 deficiency increases macrophage apoptosis and necrotic core development in atherosclerotic plaques in a Bim-dependent manner

doi: 10.1016/j.jlr.2025.100782

Figure Lengend Snippet: Deletion of myeloid Bim reduces atherosclerotic plaque, necrotic core area, and plaque apoptosis in atherosclerotic plaques of ApoA1 KO/KO Ldlr KO/KO mice. Male 10-week-old Ldlr KO/KO were transplanted with bone marrow (BM) from LyzM cre/cre ( Bim MWT ) or LyzM cre/cre Bim fl/fl ( Bim MKO ) donors and fed a high-fat diet (HFD) for 10 weeks. A: Representative images of hematoxylin and eosin (H&E) stained atherosclerotic plaques from transplanted Ldlr KO/KO mice. Dotted black line represents the necrotic core region. Analysis of B: peak plaque area and C: relative necrotic core from transplanted Ldlr KO/KO mice (n = 11, 11). Black data points represent the subset of samples used for subsequent immunofluorescent analysis. D: Representative images of TUNEL labeled (left) and CC3 stained (right) atherosclerotic plaques of transplanted Ldlr KO/KO mice. Yellow dashed line outlines the plaque area. “L” represents the lumen of the aortic valve leaflet. E: Quantification of TUNEL-positive nuclei within the atherosclerotic plaque (n = 5, 8). F: Quantification of CC3-positive area relative to the lesional area (n = 10, 11). Male 10-week-old ApoA1 KO/KO Ldlr KO/KO mice were transplanted with BM from Bim MWT or Bim MKO donors and fed a HFD for 10 weeks. G: Representative images of H&E-stained atherosclerotic plaques from transplanted ApoA1 KO/KO Ldlr KO/KO mice. Analysis of H: peak plaque area and I, relative necrotic core from transplanted ApoA1 KO/KO Ldlr KO/KO mice (n = 11, 9). J: Representative images of TUNEL labeled (left) and CC3 stained (right) atherosclerotic plaques of transplanted ApoA1 KO/KO Ldlr KO/KO mice. K: Quantification of TUNEL-positive nuclei within the atherosclerotic plaque (n = 11, 9). L: Quantification of CC3-positive area relative to the lesional area (n = 11, 9). For C–I, and L: statistical analysis was conducted using unpaired t test. For B, K: statistical analysis was conducted using Mann-Whitney test. Statistical significance is considered when P < 0.05. Data represents mean ± SEM. Bim, B-cell lymphoma 2 [Bcl-2] interacting mediator of cell death; CC3, cleaved caspase 3; LDLR, low-density lipoprotein receptor.

Article Snippet: Cells were incubated for 24 h with the absence or presence of 50 μg/ml of human HDL (J64903, Alfa Aesar, Haverhill, MA or 12-16-080412, Athens Research and Technology, Athens, GA) or 50 μg/ml of human ApoA1 (16-6-120101, Athens Research and Technology) along with differing apoptotic inducers including oxLDL (100 μg/ml, J65591, Alfa Aesar), 7-ketocholesterol (10 μg/ml; Cayman Chemical, Ann Arbor, MI), or tunicamycin (TN; 10 μg/ml; Cayman Chemical).

Techniques: Staining, TUNEL Assay, Labeling, MANN-WHITNEY

Journal: Journal of Lipid Research

Article Title: Apolipoprotein A1 deficiency increases macrophage apoptosis and necrotic core development in atherosclerotic plaques in a Bim-dependent manner

doi: 10.1016/j.jlr.2025.100782

Figure Lengend Snippet: Myeloid-specific Bim deletion in bone marrow of atherosclerotic mice does not affect plasma cholesterol levels or circulating leukocyte concentrations. Peripheral blood was collected by cardiac puncture from 10-week HFD-fed Bim MWT and Bim MKO BMT mice fasted overnight. Absolute concentrations of A: leukocytes; (B) B cells; (C) T cells; and (D) monocytes in peripheral blood samples of Ldlr KO/KO BMT mice (n = 8, 7) measured by flow cytometry. Absolute concentrations of (M) leukocytes; (N) B cells; (O) T cells; and (P) monocytes in peripheral blood samples of ApoA1 KO/KO Ldlr KO/KO BMT mice (n = 13, 10). E, Q: Ratio of Ly6C High (Hi) monocytes to Ly6C Low (Lo) monocytes in Ldlr KO/KO BMT and ApoA1 KO/KO Ldlr KO/KO BMT mice, respectively. F and R: Plasma IL-6 concentration in Ldlr KO/KO BMT (n = 12, 8) and ApoA1 KO/KO Ldlr KO/KO BMT mice (n = 8, 8), respectively. Lipids were analyzed by performing colorimetric enzymatic assays on plasma samples isolated from collected blood. Quantification of (G) total; (H) free; (I) esterified; (J) HDL, (K) non-HDL cholesterol; and (L) triglycerides from plasma of Ldlr KO/KO BMT mice (n = 12, 8). Quantification of (S) total; (T) free; (U) esterified; (V) HDL, (W) non-HDL cholesterol; and (X) triglycerides from plasma of ApoA1 KO/KO Ldlr KO/KO BMT mice (n = 13, 10). For A–H, J, M–O, R, and X: statistical analysis was conducted using unpaired t test. For I, K, L, P, Q, and S–W: statistical analysis was conducted using Mann-Whitney test. Statistical significance is considered when P < 0.05. Data represent mean ± SEM. Bim, B-cell lymphoma 2 [Bcl-2] interacting mediator of cell death; BMT, bone marrow transplantation; IL-6, interleukin-6; HFD, high-fat diet; LDLR, low-density lipoprotein receptor.

Article Snippet: Cells were incubated for 24 h with the absence or presence of 50 μg/ml of human HDL (J64903, Alfa Aesar, Haverhill, MA or 12-16-080412, Athens Research and Technology, Athens, GA) or 50 μg/ml of human ApoA1 (16-6-120101, Athens Research and Technology) along with differing apoptotic inducers including oxLDL (100 μg/ml, J65591, Alfa Aesar), 7-ketocholesterol (10 μg/ml; Cayman Chemical, Ann Arbor, MI), or tunicamycin (TN; 10 μg/ml; Cayman Chemical).

Techniques: Clinical Proteomics, Flow Cytometry, Concentration Assay, Isolation, MANN-WHITNEY, Transplantation Assay

Journal: Frontiers in Physiology

Article Title: β-Cyclodextrins Decrease Cholesterol Release and ABC-Associated Transporter Expression in Smooth Muscle Cells and Aortic Endothelial Cells

doi: 10.3389/fphys.2016.00185

Figure Lengend Snippet: Cholesterol efflux from ABAE and SMCs . Cells were first labeled with [ 3 H]-cholesterol (0.5 μCi/mL) for 36 h at 37°C and then equilibrated for 24 h in DMEM/0.05% BSA at 37°C. Cellular [ 3 H]-cholesterol loads in DPM per μg of proteins were compared between ABAE and SMCs (A) . Data represent the mean ± SEM (in DPM/μg of proteins), with n = 6 from two sets of experiments. Paired- t -test was performed, *** p < 0.001 when compared with ABAE condition. [ 3 H]-cholesterol released from both cell types was then measured 6 h after medium supplementation with either BSA (control condition) or cholesterol acceptors [ApoA-I (20 μg/mL) or HDL (50 μg/mL)] at 37°C (B) . Data represent the mean ± SEM (in DPM/μg of proteins), with n = 6 from two sets of experiments. One-way ANOVA test followed by Bonferroni correction was performed in which *** p < 0.001 refers to the control condition and ### p < 0.001 refers to the matched HDL condition in ABAE.

Article Snippet: ApoA-I and HDL were purchased from PROSCI Incorporated (Poway, CA, USA).

Techniques: Labeling, Control

Journal: Frontiers in Physiology

Article Title: β-Cyclodextrins Decrease Cholesterol Release and ABC-Associated Transporter Expression in Smooth Muscle Cells and Aortic Endothelial Cells

doi: 10.3389/fphys.2016.00185

Figure Lengend Snippet: Cholesterol release from ABAE (A,C) and from SMCs (B,D) upon CD-treatment . Cells were first labeled with [ 3 H]-cholesterol (0.5 μCi/mL) for 36 h at 37°C and then equilibrated in DMEM/0.05% BSA for 24 h at 37°C. [ 3 H]-cholesterol released in the medium from both cell types was then measured 6 h after medium supplementation with cholesterol acceptors [ApoA-I (20 μg/mL) or HDL (50 μg/mL)] at 37°C. The radioactivity contained in the culture medium, that corresponds to the cholesterol released from cells was measured. Data are expressed as the mean ± SEM (in DPM/μg of proteins, n = 6 from two sets of experiments). Statistical analysis: a one-way ANOVA followed by Dunnett's test for multiple comparisons in which * p < 0.05; *** p < 0.001 compared with the control condition (open bars) without any CD treatment.

Article Snippet: ApoA-I and HDL were purchased from PROSCI Incorporated (Poway, CA, USA).

Techniques: Labeling, Radioactivity, Control

Journal: Frontiers in Physiology

Article Title: β-Cyclodextrins Decrease Cholesterol Release and ABC-Associated Transporter Expression in Smooth Muscle Cells and Aortic Endothelial Cells

doi: 10.3389/fphys.2016.00185

Figure Lengend Snippet: Effect of 10 μM T0901317 on cholesterol efflux to ApoA-I and HDL in ABAE (A) and SMCs (B) treated or nor with Rameβ . Cells were first labeled with [ 3 H]-cholesterol as previously described and incubated in the presence of 1 mM Rameβ, 10 μM T0901317 or both during 24 h. Then, [ 3 H]-cholesterol released in the medium from both cell types was measured 6 h after medium supplementation with either ApoA-I (20 μg/mL) or HDL (50 μg/mL). Data are expressed as the mean ± SEM (in DPM/μg of proteins, n = 6 from two sets of experiments). Control condition (without any treatment) is used as the reference level and set to 100%. Statistical analysis: a one-way ANOVA followed by Dunnett's test for multiple comparisons in which * p < 0.05; ** p < 0.01; *** p < 0.001 compared with the untreated condition (Control condition, open bars) and in which ## p < 0.01 when Rameβ + T0901317 condition was compared with Rameβ condition.

Article Snippet: ApoA-I and HDL were purchased from PROSCI Incorporated (Poway, CA, USA).

Techniques: Labeling, Incubation, Control

Journal: Lipids in Health and Disease

Article Title: The mechanism of apoliprotein A1 down-regulated by Hepatitis B virus

doi: 10.1186/s12944-016-0232-5

Figure Lengend Snippet: ApoA1 expression was significantly decreased in CHB patients. a plasma ApoA1 levels were performed by ELISA in 250 CHB patients corresponding 50 healthy control. b and c ApoA1 protein levels in live tissue sections were detected by Western blot wherase ApoA1 mRNA levels from 200 CHB patients and 50 healthy controls were analyzed by RT-PCR. Results of the Real-time PCR were normalized to an endogenous control GAPDH. The ApoA1 mRNA levels in healthy controls were arbitrarily set as 1.0. Error bars are means ± standard deviation (SD). Data are presented as the mean ± SD from three independent experiments. * p < 0.05 and ** P < 0.01 compared with mock

Article Snippet: SuperscriptTM RT reagent kit (DRR037A, Takara BioInc., Shiga, Japan); ApoA1 specific siRNA and non-specific control (sc-63361, sc-37007, Santa Cruz Biotechnology); the rabbit anti-human ApoA1 (sc-30089, Santa Cruz Biotechnology); the mouse anti-human actin and the horseradish peroxidase-conjugated secondary antibodies (Zhongshan Goldenbridge Biotechnology, China); and the ECL-Plus chemiluminescence system (Applygen Technologies, Beijing, China).

Techniques: Expressing, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Control, Western Blot, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: Lipids in Health and Disease

Article Title: The mechanism of apoliprotein A1 down-regulated by Hepatitis B virus

doi: 10.1186/s12944-016-0232-5

Figure Lengend Snippet: Suppression of ApoA1 expression by HBV. a and b ApoA1 mRNA and protein levels were detected by RT-PCR and Western blot in HepG2.2.15 corresponding HepG2 cell lines. c and d HepG2 cells were transfected with 2 μg pHBV1.3 plasmid or 2 μg pCDNA3.1 as control, ApoA1 mRNA and protein levels were detected at 48 h after transfection. Data are presented as the mean ± SD from three independent experiments. * p < 0.05 and ** P < 0.01 compared with mock

Article Snippet: SuperscriptTM RT reagent kit (DRR037A, Takara BioInc., Shiga, Japan); ApoA1 specific siRNA and non-specific control (sc-63361, sc-37007, Santa Cruz Biotechnology); the rabbit anti-human ApoA1 (sc-30089, Santa Cruz Biotechnology); the mouse anti-human actin and the horseradish peroxidase-conjugated secondary antibodies (Zhongshan Goldenbridge Biotechnology, China); and the ECL-Plus chemiluminescence system (Applygen Technologies, Beijing, China).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Transfection, Plasmid Preparation, Control

Journal: Lipids in Health and Disease

Article Title: The mechanism of apoliprotein A1 down-regulated by Hepatitis B virus

doi: 10.1186/s12944-016-0232-5

Figure Lengend Snippet: ApoA1 expression was suppressed by DNA methyltransferase inhibitor 5-aza-dC. a and b 5 CpG islands including two methylation CpG status in ApoA1 promotor were listed ( a ), the detection results of the 5 CpG islands methylation status were shown ( b ). HepG2.2.15 cells treated with 5 μM 5-aza-dC 48 h, ApoA1 mRNA and protein levels were detected by RT-PCR and Western blot respectively ( c and d ). Secretion of HBsAg and HBV particles in the supernatant were detected by ELISA and RT- PCR respectivly ( e and f ). Data are presented as the mean ± SD from three independent experiments. * p < 0.05 and ** P < 0.01 compared with mock

Article Snippet: SuperscriptTM RT reagent kit (DRR037A, Takara BioInc., Shiga, Japan); ApoA1 specific siRNA and non-specific control (sc-63361, sc-37007, Santa Cruz Biotechnology); the rabbit anti-human ApoA1 (sc-30089, Santa Cruz Biotechnology); the mouse anti-human actin and the horseradish peroxidase-conjugated secondary antibodies (Zhongshan Goldenbridge Biotechnology, China); and the ECL-Plus chemiluminescence system (Applygen Technologies, Beijing, China).

Techniques: Expressing, Methylation, Reverse Transcription Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Lipids in Health and Disease

Article Title: The mechanism of apoliprotein A1 down-regulated by Hepatitis B virus

doi: 10.1186/s12944-016-0232-5

Figure Lengend Snippet: Decreased HBV expression with 5-aza-dC treatment via up-regulation of ApoA1 expression. a ApoA1 overexpression inversely suppressed HBV expression. HepG2 cells were cotransfected with 1 μg pApoA1 plasmid and 1 μg pHBV1.3. Sectetion of HBsAg and HBeAg were analyzed by ELISA at 48 h after transfection. b The inhibitory effect of 5-aza-dC on HBV expression was completely abolished by blocking 5-aza-dC-induced up-regulation of ApoA1 using RNAi. HepG2.2.15 cells were treated with 5 μM 5-aza-dC plus 50 μM ApoA1 siRNA or negative control, expression of HBsAg and HBeAg in the supernatant were analyzed by ELISA at 48 h. Data are presented as the mean ± SD from three independent experiments. * p < 0.05 and ** P < 0.01 compared with mock

Article Snippet: SuperscriptTM RT reagent kit (DRR037A, Takara BioInc., Shiga, Japan); ApoA1 specific siRNA and non-specific control (sc-63361, sc-37007, Santa Cruz Biotechnology); the rabbit anti-human ApoA1 (sc-30089, Santa Cruz Biotechnology); the mouse anti-human actin and the horseradish peroxidase-conjugated secondary antibodies (Zhongshan Goldenbridge Biotechnology, China); and the ECL-Plus chemiluminescence system (Applygen Technologies, Beijing, China).

Techniques: Expressing, Over Expression, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Transfection, Blocking Assay, Negative Control

Journal: Scientific Reports

Article Title: Comparison of the sensitivity of Western blotting between PVDF and NC membranes

doi: 10.1038/s41598-021-91521-8

Figure Lengend Snippet: Antibodies, lectins and proteins, as well as their molecular weights, transfer time and incubate time.

Article Snippet: Rabbit anti-ceruloplasmin (CerP) antibody, rabbit anti-transferrin (TF) antibody and rabbit anti-Apolipoprotein A1 (ApoA1) antibody were purchased from Boster Corporation (Wuhan, China).

Techniques: Molecular Weight, High Molecular Weight

Journal: Scientific Reports

Article Title: Comparison of the sensitivity of Western blotting between PVDF and NC membranes

doi: 10.1038/s41598-021-91521-8

Figure Lengend Snippet: Comparison of the re-probed ability of PVDF membrane and NC membrane. The pooled sera proteins (3.0, 1.5, 0.7, 0.3 and 0.1 μg) were separated by 8% SDS-PAGE, and transferred to PVDF membranes (up) and NC membrane (down), respectively. ( a ) Staining with AAL and then re-probed with PHA-E; ( b ) staining with ApoA1 and then re-probing with IgG; ( c ) Staining with PHA-E and then re-probing with A2M; ( d ) staining with A2M and then re-probing with PHA-E. Band intensities were statistically analyzed (n = 3 individual experiments) and compared using Image Lab software (Bio-Rad Laboratories) and GraphPad Prism version 6. Pink bar, PVDF membrane; Blue bar, NC membrane. Band intensities were analyzed *Significantly different p < 0.05, ** p < 0.01. N.S., not significant. All values are means ± S.E. (error bars).

Article Snippet: Rabbit anti-ceruloplasmin (CerP) antibody, rabbit anti-transferrin (TF) antibody and rabbit anti-Apolipoprotein A1 (ApoA1) antibody were purchased from Boster Corporation (Wuhan, China).

Techniques: Comparison, Membrane, SDS Page, Staining, Software

Journal: Cells

Article Title: Geniposide Ameliorated Dexamethasone-Induced Cholesterol Accumulation in Osteoblasts by Mediating the GLP-1R/ABCA1 Axis

doi: 10.3390/cells10123424

Figure Lengend Snippet: GEN ameliorated DEX-induced cholesterol accumulation by increasing ABCA1 expression. ( A , B ) The in vivo immunohistochemical examination of ABCA1 expression was conducted. ( C – E ) The protein expression of ABCA1 and ApoA-I was detected by western blot in DEX-treated MC3T3-E1 cells. ( F ) The level of the total intracellular cholesterol was measured using the ELISA kit in MC3T3-E1 cells co-treated with DEX and DIDS (200 μM). ( G – I ) The protein expression of ABCA1 and ApoA-I was detected by western blot in DEX/DIDS-treated MC3T3-E1 cells. ( J ) The ALP staining and the Alizarin Red S staining assays were conducted (×100 magnification). ( K – M ) The protein expression of RUNX2 and OPN was detected by western blot in DEX/DIDS-treated MC3T3-E1 cells. All experiments were implemented separately in triplicate. * p < 0.05; ** p < 0.01. OIM, osteogenic induction medium. NC, negative control; 50 mg/kg, Dex + 50 mg/kg GEN; 100 mg/kg, Dex + 100 mg/kg GEN.

Article Snippet: After being blocked in TBS containing 5% skimmed milk for 1 h, the membranes were co-incubated with the primary antibodies at 4 °C overnight against RUNX2 (1:1000 dilution; MyBioSource, cat.no.127554, San Diego, CA, USA), OPN (1:1000 dilution; Proteintech, cat.no.22952-1-AP, Rosemont, IL, USA), GLP-1R (1:1000 dilution; ABGENT, cat.no.AP52040), ABCA1 (1:1000 dilution; Affinity, cat.no.DF8233), apoA-I (1:1000 dilution; Affinity, cat.no.DF6264), and GAPDH (1:1000 dilution; Affinity, cat.no.AF7021), and then with HRP-labeled goat anti-rabbit secondary antibody (1:5000 dilution; Boster Biological Technology, Wuhan, China).

Techniques: Expressing, In Vivo, Immunohistochemical staining, Western Blot, Enzyme-linked Immunosorbent Assay, Staining, Negative Control

Journal: Cells

Article Title: Geniposide Ameliorated Dexamethasone-Induced Cholesterol Accumulation in Osteoblasts by Mediating the GLP-1R/ABCA1 Axis

doi: 10.3390/cells10123424

Figure Lengend Snippet: GEN promoted ABCA1-mediated cholesterol metabolism in a GLP-1R-dependent manner. ( A , B ) The in vivo immunohistochemical examination of GLP-1R expression was conducted. ( C , D ) The protein expression of GLP-1R was determined by western blot in DEX-treated MC3T3-E1 cells. ( E ) The ALP staining and the Alizarin Red S staining assay were performed in EX-treated MC3T3-E1 cells (×100 magnification). ( F ) The level of the total intracellular cholesterol was measured using the ELISA kit. ( G – K ) The protein expression of GLP-1R, ABCA1, RUNX2, and OPN was detected by western blot. All experiments were implemented separately in triplicate. * p < 0.05; ** p < 0.01. NC, negative control; 50 mg/kg, Dex + 50 mg/kg GEN; 100 mg/kg, Dex + 100 mg/kg GEN; EX, Exendin9-39.

Article Snippet: After being blocked in TBS containing 5% skimmed milk for 1 h, the membranes were co-incubated with the primary antibodies at 4 °C overnight against RUNX2 (1:1000 dilution; MyBioSource, cat.no.127554, San Diego, CA, USA), OPN (1:1000 dilution; Proteintech, cat.no.22952-1-AP, Rosemont, IL, USA), GLP-1R (1:1000 dilution; ABGENT, cat.no.AP52040), ABCA1 (1:1000 dilution; Affinity, cat.no.DF8233), apoA-I (1:1000 dilution; Affinity, cat.no.DF6264), and GAPDH (1:1000 dilution; Affinity, cat.no.AF7021), and then with HRP-labeled goat anti-rabbit secondary antibody (1:5000 dilution; Boster Biological Technology, Wuhan, China).

Techniques: In Vivo, Immunohistochemical staining, Expressing, Western Blot, Staining, Enzyme-linked Immunosorbent Assay, Negative Control