Journal: PLoS Pathogens

Article Title: Endothelial Cell Permeability during Hantavirus Infection Involves Factor XII-Dependent Increased Activation of the Kallikrein-Kinin System

doi: 10.1371/journal.ppat.1003470

Figure Lengend Snippet: Mock-, HTNV-, or ANDV-infected HUVEC were trypsinized, seeded onto ECIS chamberslides, and cultured until confluent. Media were removed and replaced with phenol red free EBM containing Zn 2+ . After an equilibration, 0.2 ml of phenol red free EBM containing Zn 2+ , FXII, PK, and HK (100 nM each) were added to cells (time zero) (black lines) (A). To measure inhibition of activation, some samples were treated with CTI (1 µM) (blue line), PKSI-527 (5 µM) (red line), or HOE 140 (1 µM) (green line) simultaneously with factors (B, C, & D). Real time measurements frequency measurements were taken throughout the assay.

Article Snippet: Arrays were then loaded into the ECIS 16-well array station and stabilized using ECIS software v1.2.92.0 (Applied Biophysics).

Techniques: Infection, Cell Culture, Inhibition, Activation Assay

Journal: FEBS Open Bio

Article Title: Cardiac pericytes function as key vasoactive cells to regulate homeostasis and disease

doi: 10.1002/2211-5463.13021

Figure Lengend Snippet: ECIS measurements of real‐time cell behavior. Quantitation of the delta max‐min of impedance to a dose response of cells to (A) PE ( n = 6, *** P = 0.0008; **** P ≤ 0.0001; P ≤ 0.0001; P ≤ 0.0001; P ≤ 0.0001, one‐way ANOVA) and (B) adenosine ( n = 6, P = 0.4751; P = 0.7496; * P = 0.0211; **** P ≤ 0.0001; P ≤ 0.0001, one‐way ANOVA). Quantitation of the delta max‐min of impedance to a dose response of cells to (C) α‐adrenergic blocker ( n = 6, P = 0.9957, P = 0.9796, **** P ≤ 0.0001, one‐way ANOVA) and (D) adenosine blockade by CGS‐15943 ( n = 6, P = 0.1370, P = 0.9997, **** P ≤ 0.0001, one‐way ANOVA). Data are presented as the mean ± SD.

Article Snippet: Inserts were placed into a Transwell filter adapter and then moved to the ECIS Ztheta 16W array module (Applied Biophysics Inc., Troy, NY, USA) inside a cell incubator at 37 °C and 5% CO 2 .

Techniques: Quantitation Assay

Journal: FEBS Open Bio

Article Title: Cardiac pericytes function as key vasoactive cells to regulate homeostasis and disease

doi: 10.1002/2211-5463.13021

Figure Lengend Snippet: Cardiac PCs were subjected to hypoxic conditions (2%) for 24 or 48 h. (A) HIF‐1α translocation after 24 h (B) Quantification of nuclear to total fluorescence of HIF‐1α protein ( n = 4, **** P ≤ 0.0001, Student's unpaired t ‐test). (C) HIF‐1α protein expression increased significantly after 24 h normalized to GAPDH ( n = 6, **** P ≤ 0.0001, Student's unpaired t ‐test). (D) Cell viability was determined over the course of 4 days. Angiogenic factors (E) PDGFbb ( n = 8, * P = 0.0005 Student's unpaired t ‐test) and VEGF‐A ( n = 8, **** P ≤ 0.0001, Student's unpaired t ‐test) were significantly increased after 48 h. (F) Apoptosis was determined by AV5 expression and PI staining where there was a significant difference between normoxic and hypoxic conditions after 48 h ( n = 3, P = 0.7802, **** P ≤ 0.0001, P ≤ 0.0001, one‐way ANOVA). ECIS measurements of real‐time cell behavior. Quantitation of the delta max‐min of impedance to a dose response of cells to (G) chemical ischemia ( n = 6, * P = 0.0144, **** P ≤ 0.0001, P ≤ 0.0001, one‐way ANOVA). Data are presented as the mean ± SD. Scale bar = 100 µm.

Article Snippet: Inserts were placed into a Transwell filter adapter and then moved to the ECIS Ztheta 16W array module (Applied Biophysics Inc., Troy, NY, USA) inside a cell incubator at 37 °C and 5% CO 2 .

Techniques: Translocation Assay, Fluorescence, Expressing, Staining, Quantitation Assay

Journal: FEBS Open Bio

Article Title: Cardiac pericytes function as key vasoactive cells to regulate homeostasis and disease

doi: 10.1002/2211-5463.13021

Figure Lengend Snippet: Effects of 24 h of LDL treatment on cardiac PCs. (A) Cell proliferation decreased upon high LDL treatment at 250 µg·mL −1 but no changes at 50 µg·mL −1 . (B) Cardiac PCs formed lipid droplets (neutral lipid staining with Oil Red O and counterstained with hematoxcylin) in response to 24 h of LDL treatment. Images taken at 10×. Black arrows point to lipid droplets in the cytoplasm at 40x after treatment at 250 µg·mL −1 . (C) Fold change in mRNA levels normalized to GAPDH. ADFP mRNA levels increased significantly upon increasing concentrations of LDL treatment ( n = 3, ** P = 0.0012, **** P < 0.0001, P < 0.0001, one‐way ANOVA). (D) Protein expression normalized to GAPDH. There is a significant increase in perilipin‐2 protein expression upon LDL treatment at 250 µg·mL −1 but not at 50 µg·mL −1 ( n = 3, P = 0.5173, **** P < 0.0001, P < 0.0001, one‐way ANOVA). ECIS measurements of real‐time cell behavior. (E) Quantitation of the delta max‐min of cell impedance to LDL treatment at 50 or 250 µg·mL −1 with or without PE‐induced contraction ( n = 6, **** P < 0.0001, two‐way ANOVA). Data are presented as the mean ± SD. Scale bar = 200 µm for 10× images. Scale bar = 100 µm for 40× image.

Article Snippet: Inserts were placed into a Transwell filter adapter and then moved to the ECIS Ztheta 16W array module (Applied Biophysics Inc., Troy, NY, USA) inside a cell incubator at 37 °C and 5% CO 2 .

Techniques: Staining, Expressing, Quantitation Assay

Journal: FEBS Open Bio

Article Title: Cardiac pericytes function as key vasoactive cells to regulate homeostasis and disease

doi: 10.1002/2211-5463.13021

Figure Lengend Snippet: Effects of 24 h of elevated d ‐glucose levels on cardiac PCs. (A) Cell proliferation was unaffected by increased glucose levels at 50 m m but cells died at low glucose levels of 5.5 m m . (B) Fold change in mRNA levels of common cytokines in diabetic conditions normalized to GAPDH. IL‐6 did not have any changes upon high glucose or high glucose in combination with insulin treatment ( n = 3, P = 0.1016, one‐way ANOVA). CCL2 increased significantly upon high glucose treatment without and with insulin ( n = 3, ** P = 0.01154 and * P = 0.02269, one‐way ANOVA). TNF‐α increased significantly upon high glucose treatment without and with insulin ( n = 3, **** P = 0.00751 and * P = 0.000738, one‐way ANOVA). (C) Protein expression of common cytokines. IL‐6 protein levels decreased significantly upon high glucose treatment with and without insulin ( n = 3, ** P = 0.0064 and * P = 0.0359, one‐way ANOVA). CCL2 protein levels did not significantly increase upon high glucose treatment but it did significantly increase upon high glucose treatment with insulin ( n = 3, P = 0.2457 and * P = 0.0472, one‐way ANOVA). ECIS measurements of real‐time cell behavior. (D) Quantitation of the delta max‐min of cell impedance to high glucose or high glucose plus insulin treatment with or without PE‐induced contraction ( n = 14, P = 0.4841, P = 0.9230, P = 0.0901, two‐way ANOVA). Data are presented as the mean ± SD.

Article Snippet: Inserts were placed into a Transwell filter adapter and then moved to the ECIS Ztheta 16W array module (Applied Biophysics Inc., Troy, NY, USA) inside a cell incubator at 37 °C and 5% CO 2 .

Techniques: Expressing, Quantitation Assay