Journal: International Journal of Nanomedicine

Article Title: Human serum albumin-based probes for molecular targeting of macrophage scavenger receptors

doi: 10.2147/IJN.S197990

Figure Lengend Snippet: Evaluation of Mal-HSA-FITC uptake by different cell types. Fluorescence microscopy images of Mal-HSA-FITC probes (green) incubated for 3 h with ( A ) THP-1 macrophages, ( C ) HUVECs (EC), and ( D ) HCtSMCs (SMC). ( B ) Mal-HSA-FITC uptake by different cell types plotted against time. The results represent data from technical triplicates (9 images/well and timepoint) and are presented as mean and SEM. Data show significantly increased uptake of modified HSA probes by THP-1 cells compared with HUVEC (**** P <0.0001) and HCtSMCs ( #### P<0.0001) at 2.5 h and 3 h. None of the cell types recognized HSA-FITC (data not shown) (repeated measures two-way ANOVA (factors: cell type and time) and Tukey’s multiple comparison test). Abbreviations: HSA, human serum albumin; GCU, green calibration unit.

Article Snippet: Human carotid smooth muscle cells (HCtSMCs) (Cell Applications, San Diego, CA, USA) were cultured up to passage 7 in human smooth muscle cell media (Cell Applications).

Techniques: Fluorescence, Microscopy, Incubation, Modification, Comparison

Journal: PLoS ONE

Article Title: Overexpression of Map3k7 activates sinoatrial node-like differentiation in mouse ES-derived cardiomyocytes

doi: 10.1371/journal.pone.0189818

Figure Lengend Snippet: Primer pairs used for real time PCR.

Article Snippet: 1 (Santa Cruz) and anti- Hcn4 (NeuroMab) antibodies.

Techniques:

Journal: PLoS ONE

Article Title: Overexpression of Map3k7 activates sinoatrial node-like differentiation in mouse ES-derived cardiomyocytes

doi: 10.1371/journal.pone.0189818

Figure Lengend Snippet: A. Immunocytochemistry showing that individual or small clusters of cardiomyocytes that form in wild-type and Map3k7 -overexpressing EBs express both the MF20 epitope (anti-CT3 epitope, green) and cardiac Troponin ( cTnI , red). Blue indicates DAPI signal in all panels. Note that Troponin protein is poorly organized in cardiac cells derived from Map3k7 -overexpressing ES cells. Scale bars indicate 10 μM in panels stained for cTnI and 20μM in panels stained for MF20. B. After several weeks in culture Map3k7 -overexpressing cells showed spider and spindle cell morphologies that are characteristic of mature SAN cells. C. Fluorescent image showing Map3k7 -overexpressing cells that also possess the MHCα::mCherry reporter (marking cardiomyocytes with red fluorescence) and wild-type cells possessing the MHCα::GFP reporter (marking cardiomyocytes with green fluorescence). Note that cardiomyocytes derived from Map3k7 -overexpressing cells possess a distinct morphology as compared to wild-type cardiomyocytes. Scale bar represents 100 μM. D. Anti- Map3k7 antibody staining of 19 day old mouse embryo showing increased expression in the remnants of the sinus venous and overlapping with HCN4 positive cells. E. By contrast, low expression of Map3k7 in the left ventricle of the same heart.

Article Snippet: 1 (Santa Cruz) and anti- Hcn4 (NeuroMab) antibodies.

Techniques: Immunocytochemistry, Derivative Assay, Staining, Fluorescence, Expressing

Journal: PLoS ONE

Article Title: Overexpression of Map3k7 activates sinoatrial node-like differentiation in mouse ES-derived cardiomyocytes

doi: 10.1371/journal.pone.0189818

Figure Lengend Snippet: A. Diagram of the major transcriptional regulators of SAN differentiation in the mouse embryo. Tbx5 activates Shox2 , Tbx3 and SAN-specific markers, including Hcn2 and Hcn4 . In addition, Shox2 inhibits Nkx2 .5 expression, and Tbx3 inhibits markers for the working myocardium. B. qRT-PCR data showing changes in the expression or timing of SAN related genes. Blue line indicates relative gene expression normalized to expression of Gapdh over time in wild-type R1 EBs and red lines indicate gene expression in Map3k7 -overexpressing EBs. Error bars indicate standard error from three technical replicates. Upregulation or downregulation of each gene was considered relevant only if relative expression trends were the same in each of a minimum of three biological replicates. Error bars represent standard error from three technical replicates. Statistical significance was determined by t-test. (*) represent p<0.006.

Article Snippet: 1 (Santa Cruz) and anti- Hcn4 (NeuroMab) antibodies.

Techniques: Expressing, Quantitative RT-PCR, Gene Expression

Journal: PLoS ONE

Article Title: Overexpression of Map3k7 activates sinoatrial node-like differentiation in mouse ES-derived cardiomyocytes

doi: 10.1371/journal.pone.0189818

Figure Lengend Snippet: A. Beat Rate Data showing quartile beat rate data for 100 beating foci for two wild type mouse strains and Map3k7 overexpressing cells. B. Immunocytochemistry showing overlap of the cardiac reporter with the SAN specific marker Hcn4 . C. Summary of this data showing the % of Hcn4 expressing cardiomyocytes is dramatically increased in the Map3k7 -overexpressing cells. D. Perforated patch recording of automaticity and pacemaker current in mCherry expressing cardiomyocytes derived from Map3k7 -overexpressing ES cells. Left: Spontaneous APs. Middle: Pacemaker current recorded from the same cell. Right: calculated activation relation from the same cell, showing current activation within the diastolic potential range. E. Calcium transients in cardiomyocytes derived from Map3k7 -overexpressing ES cells before (red trace) and after (blue trace) treatment with either 1μM norepinephrine or 0.01μM acetylcholine, as indicated. Beating in these cardiomyocytes accelerates in response to norepinephrine and slows down in response to acetylcholine.

Article Snippet: 1 (Santa Cruz) and anti- Hcn4 (NeuroMab) antibodies.

Techniques: Immunocytochemistry, Marker, Expressing, Derivative Assay, Activation Assay