red fluorescent protein mcherry  (Addgene inc)

Journal: bioRxiv

Article Title: Semaphorin 3A and 3F overexpression in TIE2 hyperactive endothelial cells contribute to the pathological lumen expansion in venous malformation

doi: 10.1101/2025.07.18.665640

Figure Lengend Snippet: (a) Volcano plot of DEGs in TIE2-L914F compared to TIE2-WT EC. A total of 860 genes showed increased expression while 1118 genes showed decreased expression. Upregulated gene expression of Sema3A and Sema3F are highlighted. (b) Gene ontology analysis of RNA sequencing of TIE2-mutant EC compared to TIE2-WT EC. In TIE2-mutant EC the following molecular functions were amongst the top 25 upregulated: Chemorepellent activity, Semaphorin receptor binding and Neuropilin binding. (c) Immmunoblot of Sema3F and Sema3A in WT-EC, TIE2-WT and TIE2-L914F EC. (d) Quantitative ELISA assay of Sema3A and (e) Sema3F protein levels in WT-EC, TIE2-WT and TIE2-L914F cells. Mean±SD, one-way ANOVA, p-values are indicated. (f) RNA Scope assay showing Sema3F (cyan) and Sema3A (magenta) expression in patient dervied tissue sections. RNA scope was combined with UEA1 fluorescent staining to identify vessels. The expression in VM lesional vessels was compared to non-lesional vessels. Scale bar: 50µm. (g) Spatial gene expression levels were quantified and expressed as % of positive signal (coverage)/nucleus. n≥100 EC nuclei/group. Mean±SD, Welch’s t-test. P-values are indicated. (h) qPCR for Sema3F and Sema3A in control EC (HUVEC and ECFC) and four VM patient-derived EC (TIE2 p.L914F mutation). Mean, quartiles, Welch’s t-test. P-values are indicated.

Article Snippet: For co-culture assays, EC were additionally transduced with lentiviral constructs encoding for either red fluorescent protein (mCherry, Addgene, cat#36084) or green fluorescent protein (GFP) (Addgene, cat#17446-LV, ).

Techniques: Expressing, Gene Expression, RNA Sequencing, Mutagenesis, Activity Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, RNAscope, Staining, Control, Derivative Assay

Journal: bioRxiv

Article Title: Modulating gene expression and protein secretion in the bacterial predator Bdellovibrio bacteriovorus

doi: 10.1101/2025.05.17.654431

Figure Lengend Snippet: ( a ) Schematic representation of B. bacteriovorus AP cell expressing cytosolic mScarletI3 under the control of P merRNA promoter on a pCAT.000-derived plasmid. ( b ) Comparison of fluorescence intensities of different fluorescent reporter proteins (mCherry, mNeonGreen, mScarletI3) expressed in B. bacteriovorus AP cells, measured at their respective emission wavelengths (610 nm, 592 nm, 517 nm respectively). The control is plasmid pCAT:P merRNA -opt.RBS lacking a fluorescent protein; its fluorescence was measured at 610 nm, 592 nm, and 517 nm, while a mean of all three measurements is shown here. White dots indicate median fluorescence. A second independent biological replicate produced highly similar results (see online source data).

Article Snippet: Coding regions for fluorescent proteins mCherry, mScarletI3 [ ], mNeonGreen [ ] were codon optimized for B. bacteriovorus (Twist Biosciences).

Techniques: Expressing, Control, Derivative Assay, Plasmid Preparation, Comparison, Fluorescence, Produced

Journal: Journal of Clinical and Translational Science

Article Title: Development of a novel single channel arteriole microphysiological system for characterizing leukocyte-endothelial interactions

doi: 10.1017/cts.2025.49

Figure Lengend Snippet: Figure 4. TNF-a induces vascular leak in SCA under flow conditions. (A) Fluorescent

Article Snippet: Both HUVEC and NHLF were transduced with lentiviruses encoding fluorescent proteins (mCherry/Addgene, 36084 or Azurite/Addgene, 36086) and were used between passages 5-9.

Techniques: