Journal: International Journal of Biological Sciences

Article Title: An apoA-I mimetic peptide facilitates off-loading cholesterol from HDL to liver cells through scavenger receptor BI

doi:

Figure Lengend Snippet: A) Induced SR-BI expression in HEK293 cells. Expression of SR-BI was induced by different concentrations of Doxycycline as indicated on the figure. Western blot analysis was performed with anti-SR-BI antibody (Novus Biologicals, NB400-113). Horseradish peroxidase-conjugated secondary antibody and the enhanced chemiluminescent substrate system (Amersham) were used. B) Bodipy-CE uptake is dependent on the expression level of SR-BI. After Doxycycline induction, HEK293[pTRE-tight-SR-BI] cells were incubated with 10 µg/mL DiO-HDL. The selective uptake was determined as described in Experimental Procedures. Data are presented as mean relative fluorescence units from triplicate or duplicate wells. Error bars represent ±SD.

Article Snippet: Western blot analysis was performed with polyclonal anti-SR-BI antibody (Novus Biologicals, NB400-113).

Techniques: Expressing, Western Blot, Incubation, Fluorescence

Journal: International Journal of Biological Sciences

Article Title: An apoA-I mimetic peptide facilitates off-loading cholesterol from HDL to liver cells through scavenger receptor BI

doi:

Figure Lengend Snippet: Bodipy-CE uptakes of reconstituted HDL and D-4F synthetic particles are mediated through SR-BI. After Doxycycline induction, HEK293[pTRE-tight-SR-BI] were incubated with 5 µg/ml Bodipy-labeled HDL and D-4F synthetic particles. The selective uptake was determined as described in "Methods". Anti-SR-BI antibody (C11, 10 µg/ml) was used in this study. Data were presented as mean relative fluorescence units from triplicate or duplicate wells. Error bars represent ±SD. The uptake of Bodipy fluorescence was blocked by SR-BI specific antibody.

Article Snippet: Western blot analysis was performed with polyclonal anti-SR-BI antibody (Novus Biologicals, NB400-113).

Techniques: Incubation, Labeling, Fluorescence

Journal: International Journal of Biological Sciences

Article Title: An apoA-I mimetic peptide facilitates off-loading cholesterol from HDL to liver cells through scavenger receptor BI

doi:

Figure Lengend Snippet: Cholesterol uptake by HepG2 cells is blocked by SR-BI specific blocking antibody C11. HepG2 cells were incubated with indicated amount of Bodipy-CE labeled particles, with or without antibody C11. The selective uptake was determined as described in Experimental Procedures. Panel A), picture taken under fluorescent microscope. HepG2 cells were incubated with 25 µg/mL of Bodipy-CE labeled rHDL in the absence of antibody. The green signal represents Bodipy; the Hoechst nuclear staining is in blue. Panel B), picture taken under fluorescent microscopic. HepG2 cells were incubated with 25 µg/mL of Bodipy-CE labeled rHDL in the presence of 10 µg/mL antibody C11. The green signal represents Bodipy; the Hoechst nuclear staining is in blue. Panel C), cholesterol uptake of Bodipy-CE of rHDL particles at different concentrations in the absence of C11 (○), in the presence of 1µg/mL C11(∆) and in the presence of 10µg/mL C11(▲ ). Panel D), cholesterol uptake of Bodipy-CE synthetic particles at 25µg/mL in the presence (empty bar) or absence of 10µg/mL C11 antibody (solid bar). Error bars represent standard deviations of relative fluorescence from triplicate determinations.

Article Snippet: Western blot analysis was performed with polyclonal anti-SR-BI antibody (Novus Biologicals, NB400-113).

Techniques: Blocking Assay, Incubation, Labeling, Microscopy, Staining, Fluorescence

Journal: Circulation

Article Title: Functional Interplay Between the Macrophage Scavenger Receptor Class B Type I and Pitavastatin (NK-104)

doi: 10.1161/01.cir.0000148368.79202.f1

Figure Lengend Snippet: Figure 1. Statins induce SR-BI expression in macrophage cell line J774 cells. J774 macrophage cell line was cultured in complete RPMI medium. At 90% confluence, cells were switched to serum-free medium and received treatments. After treatments, cells were collected to extract cellular proteins or total RNA as described in Methods. Cell lysate (20 g/sample) was used to analyze SR-BI pro- tein expression by Western blot. Total RNA (100 g) was used to isolate poly(A) RNA and detect SR-BI mRNA expression by Northern blot. A, Effect of pitavastatin on SR-BI protein expression. Macrophages were treated with indicated concentrations of pitavastatin for 16 hours. B, Effect of pitavastatin on macrophage SR-BI mRNA expression. Time of treatment was 12 hours. C, Time course of pitavastatin-induced macro-phage SR-BI protein expression. Cells were treated with 10 mol/L of pitavastatin for times indicated. D, Macrophages were treated with 10 mol/L of simvastatin (Sim), pravastatin (Pra), and rosuvastatin (Ros), respectively, for 16 hours.

Article Snippet: Rabbit polyclonal anti-human/mouse SR-BI antibody was purchased from Novus Biologicals Inc. Anti–nuclear factor (NF)- B p65 and anti–I B- antibodies were purchased from Santa Cruz Biotechnology Inc. NF- B DNA binding activity assay kits were purchased from Active-Motif, Inc.

Techniques: Expressing, Cell Culture, Western Blot, Northern Blot

Journal: Circulation

Article Title: Functional Interplay Between the Macrophage Scavenger Receptor Class B Type I and Pitavastatin (NK-104)

doi: 10.1161/01.cir.0000148368.79202.f1

Figure Lengend Snippet: Figure 3. Pitavastatin induced SR-BI expression by primary macrophages. A, Peritoneal macrophages were isolated from mice as described in Methods. Cells were cultured in completed RPMI medium and treated with indicated concentrations of pitavastatin for 16 hours. Cellular proteins were extracted, and SR-BI expression was determined by Western blot. B, Human monocytes were isolated from healthy donor’s blood and cul- tured for 1 week to differentiate macrophages. Human macro- phages were then treated with 1 and 5 mol/L of pitavastatin for 20 hours. Macrophage SR-BI surface protein was detected by FACS as described in Methods.

Article Snippet: Rabbit polyclonal anti-human/mouse SR-BI antibody was purchased from Novus Biologicals Inc. Anti–nuclear factor (NF)- B p65 and anti–I B- antibodies were purchased from Santa Cruz Biotechnology Inc. NF- B DNA binding activity assay kits were purchased from Active-Motif, Inc.

Techniques: Expressing, Isolation, Cell Culture, Western Blot

Journal: Circulation

Article Title: Functional Interplay Between the Macrophage Scavenger Receptor Class B Type I and Pitavastatin (NK-104)

doi: 10.1161/01.cir.0000148368.79202.f1

Figure Lengend Snippet: Figure 4. Pitavastatin restored macrophage SR-BI expression after inhibition by cholesterol biosynthetic intermediates. Conflu- ent J774 macrophages were treated with mevalonate (10 and 50 mol/L, A), farnesyl-pyrophosphate (FPP, 1 and 5 mol/L, B), geranylgeranyl-pyrophosphate (GG-PP, 1 and 5 mol/L, C), and coenzyme Q10 (CoQ10, 2 and 5 mol/L, D), respectively, in presence and absence of pitavastatin (10 mol/L) for 16 hours in serum-free medium. Cell lysate was extracted and analyzed for SR-BI expression by Western blot.

Article Snippet: Rabbit polyclonal anti-human/mouse SR-BI antibody was purchased from Novus Biologicals Inc. Anti–nuclear factor (NF)- B p65 and anti–I B- antibodies were purchased from Santa Cruz Biotechnology Inc. NF- B DNA binding activity assay kits were purchased from Active-Motif, Inc.

Techniques: Expressing, Inhibition, Western Blot

Journal: Circulation

Article Title: Functional Interplay Between the Macrophage Scavenger Receptor Class B Type I and Pitavastatin (NK-104)

doi: 10.1161/01.cir.0000148368.79202.f1

Figure Lengend Snippet: Figure 5. NF-B inhibitors blocked inhibition of macrophage SR-BI expression by LPS and TNF-. A, Macrophages were treated with LPS (200 ng/mL) or NF-B inhibitor (Bay 11-7083 and Bay 11-7085, 5 mol/L in each) or LPS plus NF-B inhibitor for 16 hours. Cell lysate was extracted, and SR-BI expression was determined by Western blot. B, Macrophages were treated with TNF- or NF-B inhibitor (Bay 11-7083, Bay 11-7085, NF-B SN50) or TNF- plus NF-B inhibitor at indicated con- centration for 16 hours. Cell lysate was used to assess macro- phage SR-BI expression by Western blot.

Article Snippet: Rabbit polyclonal anti-human/mouse SR-BI antibody was purchased from Novus Biologicals Inc. Anti–nuclear factor (NF)- B p65 and anti–I B- antibodies were purchased from Santa Cruz Biotechnology Inc. NF- B DNA binding activity assay kits were purchased from Active-Motif, Inc.

Techniques: Inhibition, Expressing, Western Blot

Journal: Circulation

Article Title: Functional Interplay Between the Macrophage Scavenger Receptor Class B Type I and Pitavastatin (NK-104)

doi: 10.1161/01.cir.0000148368.79202.f1

Figure Lengend Snippet: Figure 6. Pitavastatin restored macrophage SR-BI expression after inhibition by LPS and TNF-. Confluent macrophages in serum-free medium were treated with various concentrations of LPS (A) or TNF- (B) in presence or absence of 5 mol/L pitavastatin for 16 hours. Cell lysate was extracted and was then used to assess SR-BI expression by Western blot.

Article Snippet: Rabbit polyclonal anti-human/mouse SR-BI antibody was purchased from Novus Biologicals Inc. Anti–nuclear factor (NF)- B p65 and anti–I B- antibodies were purchased from Santa Cruz Biotechnology Inc. NF- B DNA binding activity assay kits were purchased from Active-Motif, Inc.

Techniques: Expressing, Inhibition, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Low-Density Lipoprotein Receptor (LDLR) Is Involved in Internalization of Lentiviral Particles Pseudotyped with SARS-CoV-2 Spike Protein in Ocular Cells

doi: 10.3390/ijms241411860

Figure Lengend Snippet: Antibodies used in experiments.

Article Snippet: SR-B1 rabbit polyclonal IgG , NB400-113 , Novus Biologicals (Centennial, CO, USA).

Techniques: Purification