Additional file 2 of Sex-dependent expression of neutrophil gelatinase-associated lipocalin in aortic stenosis
Creators: name: Jover, Eva
name: Matilla, Lara
name: Martín-Núñez, Ernesto
name: Garaikoetxea, Mattie
name: Navarro, Adela
name: Fernández-Celis, Amaya
name: Gainza, Alicia
name: Arrieta, Vanessa
name: García-Peña, Amaia
name: Álvarez, Virginia
name: Sádaba, Rafael
name: Jaisser, Frederic
name: López-Andrés, Natalia
Publisher: name: figshare
Year: 2023
Subjects: subject: Biochemistry
subject: Medicine
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Physiology
subject: 39999 Chemical Sciences not elsewhere classified
subject: FOS: Chemical sciences
subject: Immunology
subject: FOS: Clinical medicine
subject: 69999 Biological Sciences not elsewhere classified
subject: Cancer
subject: 110309 Infectious Diseases
subject: FOS: Health sciences

description: Additional file 2: Figure S1. Effect of HP in VICs isolated from men and women donors. Scatter dot plot of calcium deposits in primary cultured VICs challenged with HP (2.6mM Pi) (A). Representative immunoblots for intracellular NGAL expression in male and female-derived VICs (D). **** p &lt; 0.0001. Ctrl, control; NGAL, Neutrophil Gelatinase-Associated Lipocalin. Figure S2. Effect of NGAL silencing on control VICs from men donors undergoing elective surgical AV replacement. NGAL silencing was evidenced at transcript and protein levels in control conditions (A &amp; B). Scatter dot plots for the relative expression of NGAL codifying gene (LCN2) in control male-derived VICs (A). Scramble control group was used as calibrator for the fold-change calculations. GAPDH, 18S, HPRT and ACTA2 were used as housekeeping genes. Representative immunoblots for NGAL expression in man-derived VICs (B). Protein lysates were harvested at day 4 and were assayed into native SDS-PAGE. Protein quantifications were normalized to Stain free. Markers of inflammation (C), ECM remodelling (D) and osteogenesis (E) were assessed by ELISA in cell supernatants from siNGAL VICs under control conditions. Osteogenic markers RUNX2 and BMP2 were analysed at the mRNA transcript level as well as the quiescent VIC marker LECT1 (chondromodulin-I) (F). AU, arbitrary units; *, p &lt; 0.05; **, p &lt; 0.01; ***, p &lt; 0.001; p &lt; 0.0001. LCN2, NGAL codifying gene; CCL2, chemokine (C-C motif) ligand 2 (CCL2) or monocyte chemoattractant protein 1 (MCP1); MPO, myeloperoxidase; Gal-3, galectin-3; MMP, metalloproteinase; TIMP, tissue inhibitor of metalloproteinases; BMP, bone morphogenetic protein; LECT1, chondromodulin (Chm)-I codifying gene;, periostin; RUNX2, Runt-related transcription factor 2. Figure S3. Effect of exogenous NGAL regulation in control VICs isolated from men donors. Markers of inflammation (A), ECM remodelling (B) and osteogenesis (C) were assessed by ELISA in cell supernatants from male-derived VICs silenced for NGAL and under control conditions. All comparisons have been represented as a fold change relative to Control condition. **, p &lt; 0.01;. CCL2, chemokine (C-C motif) ligand 2 (CCL2) or monocyte chemoattractant protein 1 (MCP1); MPO, myeloperoxidase; Gal-3, galectin-3; TNF-a, tumour necrosis factor-a; IL, interleukin; MMP, metalloproteinase; TIMP, tissue inhibitor of metalloproteinases; BMP, bone morphogenetic protein.