Journal: Circulation Research

Article Title: Loss of Plasma Proprotein Convertase Subtilisin/Kexin 9 (PCSK9) After Lipoprotein Apheresis

doi: 10.1161/circresaha.113.302655

Figure Lengend Snippet: Figure 1. Lipoprotein apheresis (LA) reduces plasma proprotein convertase subtilisin/kexin 9 (PCSK9) levels. A, Pre- and postapheresis plasma PCSK9 levels in 6 patients with familial hypercholesterolemia (FH). Insert: Representative PCSK9 immunoblot from pre- and post-apheresis plasma. B, Pre- and post-apheresis plasma PCSK9 levels during 3 consecutive treatments in 6 FH patients. C, Correlation between low-density lipoprotein (LDL) reduction and PCSK9 reduction after LA. ***P<0.001.

Article Snippet: Sheep polyclonal antibody toward human PCSK9 was purchased from R&D systems (Minneapolis, MN).

Techniques: Clinical Proteomics, Western Blot

Journal: Circulation Research

Article Title: Loss of Plasma Proprotein Convertase Subtilisin/Kexin 9 (PCSK9) After Lipoprotein Apheresis

doi: 10.1161/circresaha.113.302655

Figure Lengend Snippet: Figure 3. Cholesterol and proprotein convertase subtilisin/kexin 9 (PCSK9) adsorption to a scaled-down dextran sulfate column. A, Percent of cholesterol removed from total plasma and its low-density lipoprotein (LDL) and apoB-free fractions by a scaled-down dextran sulfate column (n=3). B, Percent of PCSK9 removed from plasma, its fractions, and purified GST-tagged PCSK9 by the scaled- down dextran sulfate column (n=3). Insert: PCSK9 immunoblot of GST-tagged eluted fraction from glutathione agarose column of media from HEK293T cells transfected with pcDNA-PCSK9-GST. The presence of a single band confirms the purity of the preparation.

Article Snippet: Sheep polyclonal antibody toward human PCSK9 was purchased from R&D systems (Minneapolis, MN).

Techniques: Adsorption, Clinical Proteomics, Purification, Western Blot, Transfection

Journal: Molecular Medicine Reports

Article Title: Effects of Tanshinone IIA on the modulation of miR-33a and the SREBP-2/Pcsk9 signaling pathway in hyperlipidemic rats

doi: 10.3892/mmr.2016.5133

Figure Lengend Snippet: Tanshinone IIA regulation of SREBP-2, Pcsk9 and LDL-R mRNA and protein expression levels in the liver. (A) Reverse transcription-polymerase chain reaction measurement of SREBP-2, Pcsk9 and LDL-R mRNA expression levels in rat liver tissue. (B) Western blotting analysis of SREBP-2, Pcsk9 and LDL-R protein expression levels in the liver. Data are presented as the mean ± standard deviation. * P<0.05, ** P<0.01. SREBP-2, sterol regulatory element-binding protein 2; Pcsk9, proprotein convertase subtilisin/kexin type 9; LDL-R, low-density lipoprotein receptor; CON, control; HLP, high lipid diet-fed rats; TAN, Tanshinone IIA treatment of high lipid diet-fed rats; GAPDH, .glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: Subsequently, the sections were incubated with primary polyclonal rabbit anti-rat SREBP-2 (cat no. 980594w), LDL-R (cat no. YSLS15w), PCSK9 (cat no. bs-6060R) antibodies (all from BIOSS, Beijing, China). at 4°C overnight, then washed with PBS three times and incubated with a horseradish peroxidase (HRP)-conjugated goat anti-rabbit secondary antibody (1:1,000; 36J00180; Beijing Dingguo Changsheng Biotechnology Co., Ltd., Beijing, China) at room temperature for 1 h. The peroxidase activity was detected using 3, 3′-diaminobenzidine tetrahydrochloride solution.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Standard Deviation, Binding Assay

Journal: Molecular Medicine Reports

Article Title: Effects of Tanshinone IIA on the modulation of miR-33a and the SREBP-2/Pcsk9 signaling pathway in hyperlipidemic rats

doi: 10.3892/mmr.2016.5133

Figure Lengend Snippet: Immunohistochemical detection of SREBP-2, Pcsk9 and LDL-R expression in the liver tissue. Tissue from the CON, HLP and TAN groups was collected and stained with antibodies against SREBP-2, Pcsk9 and LDL-R. (A) SREBP-2 protein expression in liver tissue. (B) Pcsk9 protein expression in liver tissue. Data are presented as the mean ± standard deviation. ** P<0.01. SREBP-2, sterol regulatory element-binding protein 2; Pcsk9, proprotein convertase subtilisin/kexin type 9; LDL-R, low-density lipoprotein receptor; CON, control; HLP, high lipid diet-fed rats; TAN, Tanshinone IIA treatment of high lipid diet-fed rats; IOD, integrated optical density. (C) LDL-R protein expression in liver tissue. Data are presented as the mean ± standard deviation. ** P<0.01. LDL-R, low-density lipoprotein receptor; CON, control; HLP, high lipid diet-fed rats; TAN, Tanshinone IIA treatment of high lipid diet-fed rats; IOD, integrated optical density.

Article Snippet: Subsequently, the sections were incubated with primary polyclonal rabbit anti-rat SREBP-2 (cat no. 980594w), LDL-R (cat no. YSLS15w), PCSK9 (cat no. bs-6060R) antibodies (all from BIOSS, Beijing, China). at 4°C overnight, then washed with PBS three times and incubated with a horseradish peroxidase (HRP)-conjugated goat anti-rabbit secondary antibody (1:1,000; 36J00180; Beijing Dingguo Changsheng Biotechnology Co., Ltd., Beijing, China) at room temperature for 1 h. The peroxidase activity was detected using 3, 3′-diaminobenzidine tetrahydrochloride solution.

Techniques: Immunohistochemical staining, Expressing, Staining, Standard Deviation, Binding Assay

Journal: Cell Death & Disease

Article Title: Reciprocal positive regulation between Cx26 and PI3K/Akt pathway confers acquired gefitinib resistance in NSCLC cells via GJIC-independent induction of EMT

doi: 10.1038/cddis.2015.197

Figure Lengend Snippet: Increased Cx26 is positively correlated with gefitinib resistance in NSCLC cells. ( a ) Differential expression of Cx26, Cx31.1, Cx32, and Cx43 in different gefitinib-sensitive NSCLC cell lines was determined by RT-PCR. ( b and c ) High level of Cx26 in gefitinib-insensitive A549 and H1299 cells than that in gefitinib-sensitive HCC827 and PC9 cells was detected by RT-PCR and western blotting. GAPDH or β -actin was used as internal loading control

Article Snippet: Human NSCLC cell lines (HCC827, PC9, A549, and H1299) were originally obtained from the ATCC (Manassas, VA, USA).

Techniques: Quantitative Proteomics, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control

Journal: Cell Death & Disease

Article Title: Reciprocal positive regulation between Cx26 and PI3K/Akt pathway confers acquired gefitinib resistance in NSCLC cells via GJIC-independent induction of EMT

doi: 10.1038/cddis.2015.197

Figure Lengend Snippet: Cx26 induces acquired gefitinib resistance in NSCLC cells via GJIC-independent manner. ( a ) Functional GJIC was detected by parachute assay and no detectable GJIC was found in HCC827 GR, PC9 GR, and their parental cells. Top: fluorescence images. Bottom: overlaid the corresponding phase-contrast images. Original magnification, × 200. ( b ) No enhancement of GJIC in these cells incubated with 10, 20, and 40 μ M of RA (a well-defined GJIC enhancer) for 4, 8, 12, 24, and 48 h, respectively. Top: fluorescence images. Bottom: overlaid the corresponding phase-contrast images. Original magnification, × 200. ( c and d ) Immunofluorescence staining of the cellular localization of Cx26 with or without RA treatment. All scare bars represent 50 μ m

Article Snippet: Human NSCLC cell lines (HCC827, PC9, A549, and H1299) were originally obtained from the ATCC (Manassas, VA, USA).

Techniques: Functional Assay, Fluorescence, Incubation, Immunofluorescence, Staining

Journal: Cell Death & Disease

Article Title: Reciprocal positive regulation between Cx26 and PI3K/Akt pathway confers acquired gefitinib resistance in NSCLC cells via GJIC-independent induction of EMT

doi: 10.1038/cddis.2015.197

Figure Lengend Snippet: Cx26 and PI3K/Akt pathway functionally interplay to promote EMT and gefitinib resistance in NSCLC cells. ( a and b ) Effect of LY294002 or Akt overexpression on Cx26 expression in HCC827, PC9, and their GR cells was determined by western blotting. ( c ) Effects of Akt overexpression alone or combined with Cx26 overexpression or Cx26 depletion on cell morphology changes in HCC827 and PC9 cells. Original magnification, × 400. ( d – f ) Effects of Akt overexpression alone or combined with Cx26 overexpression or Cx26 depletion on the expression of EMT markers (E-cadherin, vimentin, and slug), cell migration, and invasion, as well as cell sensitivity to gefitinib in HCC827 and PC9 cells, respectively. Error bars are mean±S.D. from four independent experiments, ** P <0.01 versus vector group. # P <0.05 and ## P <0.01 versus Akt-overexpressing group

Article Snippet: Human NSCLC cell lines (HCC827, PC9, A549, and H1299) were originally obtained from the ATCC (Manassas, VA, USA).

Techniques: Over Expression, Expressing, Western Blot, Migration, Plasmid Preparation

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Ser-Phosphorylation of PCSK9 by Fam20C-Kinase Enhances its Ability to Degrade the LDLR

doi: 10.1161/ATVBAHA.119.313247

Figure Lengend Snippet: Comparative mRNA distribution of mouse Fam20A, Pcsk9 and Fam20C.

Article Snippet: PCSK9-LDLR binding affinity assay The PCSK9-LDLR binding affinity was assessed by using a CircuLex human PCSK9 functional assay kit (MBL, Cat # CY8153) following the manufacturers’ instructions.

Techniques:

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Ser-Phosphorylation of PCSK9 by Fam20C-Kinase Enhances its Ability to Degrade the LDLR

doi: 10.1161/ATVBAHA.119.313247

Figure Lengend Snippet: Effects of PCSK9-phosphosphomimetics on LDLR degradation.

Article Snippet: PCSK9-LDLR binding affinity assay The PCSK9-LDLR binding affinity was assessed by using a CircuLex human PCSK9 functional assay kit (MBL, Cat # CY8153) following the manufacturers’ instructions.

Techniques:

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Ser-Phosphorylation of PCSK9 by Fam20C-Kinase Enhances its Ability to Degrade the LDLR

doi: 10.1161/ATVBAHA.119.313247

Figure Lengend Snippet: Fam20C/Fam20A enhance PCSK9 secretion and activity on the LDLR.

Article Snippet: PCSK9-LDLR binding affinity assay The PCSK9-LDLR binding affinity was assessed by using a CircuLex human PCSK9 functional assay kit (MBL, Cat # CY8153) following the manufacturers’ instructions.

Techniques: Activity Assay

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Ser-Phosphorylation of PCSK9 by Fam20C-Kinase Enhances its Ability to Degrade the LDLR

doi: 10.1161/ATVBAHA.119.313247

Figure Lengend Snippet: In vitro and cellular validation of Fam20C-targets on PCSK9.

Article Snippet: PCSK9-LDLR binding affinity assay The PCSK9-LDLR binding affinity was assessed by using a CircuLex human PCSK9 functional assay kit (MBL, Cat # CY8153) following the manufacturers’ instructions.

Techniques: In Vitro

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Ser-Phosphorylation of PCSK9 by Fam20C-Kinase Enhances its Ability to Degrade the LDLR

doi: 10.1161/ATVBAHA.119.313247

Figure Lengend Snippet: Characterization of Fam20C-mediated Ser-phosphorylation on PCSK9 activity.

Article Snippet: PCSK9-LDLR binding affinity assay The PCSK9-LDLR binding affinity was assessed by using a CircuLex human PCSK9 functional assay kit (MBL, Cat # CY8153) following the manufacturers’ instructions.

Techniques: Activity Assay

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Ser-Phosphorylation of PCSK9 by Fam20C-Kinase Enhances its Ability to Degrade the LDLR

doi: 10.1161/ATVBAHA.119.313247

Figure Lengend Snippet: Effect of PCSK9 pro-domain and Furin in PCSK9-mediated LDLR degradation.

Article Snippet: PCSK9-LDLR binding affinity assay The PCSK9-LDLR binding affinity was assessed by using a CircuLex human PCSK9 functional assay kit (MBL, Cat # CY8153) following the manufacturers’ instructions.

Techniques: