Journal: International Journal of Molecular Sciences

Article Title: Human ApoA-I Overexpression Enhances Macrophage-Specific Reverse Cholesterol Transport but Fails to Prevent Inherited Diabesity in Mice

doi: 10.3390/ijms20030655

Figure Lengend Snippet: Hepatic and intestinal gene expression profiles of molecular targets involved in macrophage-specific reverse cholesterol transport (m-RCT).

Article Snippet: Specific mouse Taqman probes (Applied Biosystems) were used for Apoa1 (Mm00437569_m1), Abca1 (Mm00442646_m1), Abcg1 (Mm00437390_m1), Abcg5 (Mm00446241_m1), Abcg8 (Mm00445970_m1), Abcb11 (Mm00446241_m1), Acaca (Mm01304257_m1), Cd36 (Mm01135198_m1), Cyp7a1 (Mm00484152_m1), Cyp27a1 (Mm00470430_m1), Hmgcr (Mm01282499_m1), Hspa5 (Mm00517691_m1), Ncp1l1 (Mm01191972_m1), Scarb1 (Mm00450236_m1), and Gapdh (Mm99999915_g1) (reference gene) to analyze gene expression in mouse tissues.

Techniques: Gene Expression

Journal: International Journal of Molecular Sciences

Article Title: Differences in Functional Expression of Connexin43 and Na V 1.5 by Pan- and Class-Selective Histone Deacetylase Inhibition in Heart

doi: 10.3390/ijms19082288

Figure Lengend Snippet: Changes in cardiac Cx43 and Na V 1.5 protein levels by panobinostat, entinostat and ricolinostat. ( A ) Representative Western blot of lysed NMVMs treated with 0, 25, 100, 500, and 2500 nM panobinostat for 24 h. Ac-tubulin is acetylated α-tubulin, Ac-H3 is acetylated histone 3, and α-tubulin was used as a loading control. ( B ) Densitometry scans of Cx43 Western blots ( n = 3) to quantify the reduction in Cx43 protein levels with increasing concentrations of panobinostat (* p < 0.05). ( C ) Densitometry scans of Na V 1.5 Western blots ( n = 3) quantifying the statistically significant decrease in Na V 1.5 protein levels with higher concentrations of panobinostat (* p < 0.05). ( D ) A representative Western blot of lysed ventricular myocytes treated with 1 μM entinostat (MS-275) or 25 nM ricolinostat for 24 h. Note that MS-275 only increased the Ac-H3 signal while ACY-1215 only increased the Ac-α-tubulin signal, consistent with their class I and HDAC6 inhibitory activities. ( E ) Densitometry scans of 1 μM entinostat Western blots ( n = 4) illustrating no significant changes in Cx43 or Na V 1.5 protein levels with entinostat, a class I HDAC-selective inhibitor. ( F ) Densitometry scans of 25 nM ricolinostat Western blots ( n = 3) illustrating no significant changes in Cx43 or Na V 1.5 protein levels with ricolinostat.

Article Snippet: Primary antibodies used in this study include rabbit anti-Cx43 (AB1728, Merck Millipore, Billerica, MA, USA), mouse anti-Cx43 (AB1727, Merck Millipore), mouse anti-α-tubulin (# T5168, Sigma-Aldrich, St. Louis, MO, USA), rabbit anti-acetylated H3 (# 06-599, Merck Millipore), rabbit anti-acetylated-α-tubulin (BML-SA452-0100, Enzo Life Sciences, Farmingdale, NY, USA) and rabbit anti-Na V 1.5 (# ACC-001, Alomone Labs, Jerusalem, Israel).

Techniques: 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

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