Journal: Renal Failure

Article Title: PCSK6 is a novel regulator of venous smooth muscle cell function in arteriovenous fistula remodeling

doi: 10.1080/0886022X.2026.2663246

Figure Lengend Snippet: PCSK6 is increased in smooth muscle cells of stenotic arteriovenous fistula. (A-B) Human stenotic and non-stenotic arteriovenous fistula (AVF) tissues were obtained as described in the Materials and Methods. (A) Representative images of hematoxylin and eosin (HE) staining and immunofluorescence staining for PCSK6, COL1A1, and MYH11 in tissue sections. Immunofluorescence intensity of PCSK6 in the two groups, as well as correlations between PCSK6 and COL1A1 immunofluorescence intensity, neointimal thickness, degree of luminal stenosis, and AVF blood flow were plotted. (B) Total protein and RNA were extracted from tissues. Protein expression of PCSK6 was analyzed by Western blot, and mRNA expression was determined by real-time PCR. (C-D) Primary cultured smooth muscle cells (SMCs) were derived from human stenotic and non-stenotic AVF tissues. (C) Immunofluorescence staining for PCSK6 and the SMC marker MYH11 in primary cultured SMCs. (D) Total protein and RNA were extracted from primary cultured SMCs. Protein expression of PCSK6 was analyzed by Western blot, and mRNA expression was determined by real-time PCR.

Article Snippet: Human venous smooth muscle cells were purchased from ATCC with the Vascular Smooth Muscle Cell Growth Kit (ATCC PCS-100-04).

Techniques: Staining, Immunofluorescence, Expressing, Western Blot, Real-time Polymerase Chain Reaction, Cell Culture, Derivative Assay, Marker

Journal: Renal Failure

Article Title: PCSK6 is a novel regulator of venous smooth muscle cell function in arteriovenous fistula remodeling

doi: 10.1080/0886022X.2026.2663246

Figure Lengend Snippet: PCSK6 is increased in smooth muscle cells during venous remodeling after arteriovenous creation. (A-B) Mouse AVF models were generated as described in the Material and Methods. (A) Tissues from the AVF anastomosis were collected at the indicated time points. Representative images of HE staining and immunofluorescence staining for PCSK6 and MYH11 are shown. Neointimal thickness and PCSK6 immunofluorescence intensity across different time points, as well as the correlation between PCSK6 intensity and neointimal thickness were plotted. (B) Total protein and RNA were extracted from the tissues. Protein expression of PCSK6 at different time points was analyzed by Western blot, and mRNA expression was determined by real-time PCR. (C-D) Primary cultured SMCs were derived from the AVF at the indicated time points. (C) Immunofluorescence staining for PCSK6 and the SMC marker MYH11 in primary cultured SMCs. (D) Total protein and RNA were extracted from primary cultured SMCs. Protein expression of PCSK6 was analyzed by Western blot, and mRNA expression was determined by real-time PCR.

Article Snippet: Human venous smooth muscle cells were purchased from ATCC with the Vascular Smooth Muscle Cell Growth Kit (ATCC PCS-100-04).

Techniques: Generated, Staining, Immunofluorescence, Expressing, Western Blot, Real-time Polymerase Chain Reaction, Cell Culture, Derivative Assay, Marker

Journal: Renal Failure

Article Title: PCSK6 is a novel regulator of venous smooth muscle cell function in arteriovenous fistula remodeling

doi: 10.1080/0886022X.2026.2663246

Figure Lengend Snippet: PCSK6 promotes smooth muscle cells phenotypic switch and ECM production. (A–G) Venous SMCs were transfected with control or PCSK6 expression vectors. (A) Total protein and RNA were extracted. Protein expression of COL1A1, fibronectin, VIM, and MMP2 was analyzed by Western blot, and mRNA expression was determined by real-time PCR. (B) Cell viability was assessed using CCK-8 assay. (C) Cell proliferation was measured by BrdU assay. (D) Cell migration was evaluated by wound healing assay. (E) Cell contractility was determined by collagen gel contraction assay. (F). Hydroxyproline levels were quantified. (G) MMPs activity was measured using MMPs activity kit as described in the Material and Methods section. (H–N) PrimaryM cultured SMCs were transfected with siRNA targeting either control or PCSK6. (H) Total protein and RNA were extracted. Protein expression of COL1A1, fibronectin, VIM, and MMP2 was analyzed by Western blot, and mRNA expression was determined by real-time PCR. (I) Cell viability was assessed using CCK-8 assay. (J) Cell proliferation was measured by BrdU assay. (K) Cell migration was evaluated by wound healing assay. (L) Cell contractility was determined by collagen gel contraction assay. (M) Hydroxyproline levels were quantified. (N) MMPs activity was measured using MMPs activity kit as described in the Material and Methods section.

Article Snippet: Human venous smooth muscle cells were purchased from ATCC with the Vascular Smooth Muscle Cell Growth Kit (ATCC PCS-100-04).

Techniques: Transfection, Control, Expressing, Western Blot, Real-time Polymerase Chain Reaction, CCK-8 Assay, BrdU Staining, Migration, Wound Healing Assay, Collagen Gel Contraction Assay, Activity Assay, Cell Culture

Journal: Renal Failure

Article Title: PCSK6 is a novel regulator of venous smooth muscle cell function in arteriovenous fistula remodeling

doi: 10.1080/0886022X.2026.2663246

Figure Lengend Snippet: Silencing of PCSK6 in VSMCs alleviated venous remodeling and AVF stenosis. (A) Smooth muscle cell-specific PCSK6 knockout mice were generated as described in the Material and Methods. The schematic illustrates the experimental timeline after AVF creation in both knockout and control mice. (B) AVF diameter and blood flow were monitored by ultrasound. Quantitative data are presented. (C-D) Functional analysis of harvested IVC segments assessing (C) contraction responses to 40mM KCl and (D) Relaxation responses to the cumulative addition of acetylcholine. (E) Total protein and RNA were extracted. Protein expression of COL1A1, fibronectin, MMP2 and VIM, was analyzed by Western blot, and mRNA expression was determined by real-time PCR. (F) Histological evaluation of IVC sections through HE/EVG/Masson staining and immunofluorescence for PCSK6 and MYH11. Neointimal thickness was quantified in both experimental groups.

Article Snippet: Human venous smooth muscle cells were purchased from ATCC with the Vascular Smooth Muscle Cell Growth Kit (ATCC PCS-100-04).

Techniques: Knock-Out, Generated, Control, Functional Assay, Expressing, Western Blot, Real-time Polymerase Chain Reaction, Staining, Immunofluorescence