human aorta smcs  (Kurabo industries)

Journal: International Journal of Molecular Sciences

Article Title: Extracellular Matrix Analysis of Human Renal Arteries in Both Quiescent and Active Vascular State

doi: 10.3390/ijms21113905

Figure Lengend Snippet: ( A ) Representative images demonstrate the distribution of EMILIN1 co-stained with smooth muscle actin (αSMA) in fetal and mature human renal arteries. Co-localization of αSMA and EMILIN1 in yellow. ( B ) Representative images demonstrate the distribution of FBN1 co-stained with αSMA in fetal and mature human renal arteries. Co-localization of αSMA and FBN1 in yellow. Scale bar represents 50 µm. Open lumen is indicated with a cross, tunica media layer is indicated with an asterix, and the tunica adventitia layer is indicated with a pound sign. ( C ) Quantification of EMILIN1 and αSMA positive signal in percentages in fetal and mature renal arteries. ( D ) Quantification of FBN1 and αSMA positive signal in percentages in fetal and mature renal arteries. Data are shown as mean ± SEM, N = 4–5 fluorescent images for EMILIN1 and FBN1 respectively, in fetal samples. N = 11–15 fluorescent images for EMILIN1 and FBN1 respectively, in mature samples. * p < 0.05, **** p < 0.0001 (Student’s t -test).

Article Snippet: Human aorta smooth muscle cells (SMCs; Lonza) were cultured on gelatin-coated plates in smooth muscle growth medium (SMBM basal medium supplemented with SMGM bullet kit; Lonza) in 5% CO 2 at 37 °C.

Techniques: Staining

Journal: International Journal of Molecular Sciences

Article Title: Extracellular Matrix Analysis of Human Renal Arteries in Both Quiescent and Active Vascular State

doi: 10.3390/ijms21113905

Figure Lengend Snippet: ( A ) qPCR analysis of EMILIN1 and FBN1 in different cell types of the vasculature, human umbilical vein endothelial cells (HUVECs; as endothelium source), pericytes and vascular smooth muscle cells (vSMC; hereafter referred as SMC). Data are shown as mean ± SEM corrected for beta-actin (housekeeping gene). N ≤ 10, * p < 0.05, ** p < 0.01, **** p < 0.0001 compared to HUVECs, # p < 0.05 compared to pericytes (One-way analysis of variance (ANOVA), Tukey’s post hoc test). ( B ) qPCR validation of EMILIN1 and FBN1 knockdown in SMC 6 days after siRNA transfection. Data are shown as mean ± SEM, N = 7–9 for EMILIN1 and FBN1 , respectively. ** p < 0.01, *** p < 0.001 compared to siSham (Student’s t -test). ( C ) Representative z-stacks of EMILIN1 on SMC cultured for 6 days after siRNA transfection and after decellularization. Scale bar represents 100 µm. ( D ) Quantification of EMILIN1 signal-corrected for the amount of F-actin in siRNA-treated SMC. Data are shown as mean ± SEM, N = 5. ( E ) Quantification of EMILIN1 signal-corrected for the amount of collagen type IV present on SMC-derived ECM coverslips. Data are shown as mean ± SEM, N = 7, ** p < 0.01, *** p < 0.0001 (Student’s t -test). Non-treated SMC are set to one (dotted lines).

Article Snippet: Human aorta smooth muscle cells (SMCs; Lonza) were cultured on gelatin-coated plates in smooth muscle growth medium (SMBM basal medium supplemented with SMGM bullet kit; Lonza) in 5% CO 2 at 37 °C.

Techniques: Biomarker Discovery, Knockdown, Transfection, Cell Culture, Derivative Assay