Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Evaluation of suitable reference genes for normalization of real-time reverse transcription PCR analysis in colon cancer

doi: 10.1186/1756-9966-29-144

Figure Lengend Snippet: Candidate reference genes included in the TaqMan Endogenous Control Assay.

Article Snippet: Transferrin receptor (p90, CD71) , TFRC , Hs99999911_m1 , 105 , Cellular uptake of iron.

Techniques: Control, Immunopeptidomics, Binding Assay, Ubiquitin Proteomics, Activation Assay, Transduction

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Evaluation of suitable reference genes for normalization of real-time reverse transcription PCR analysis in colon cancer

doi: 10.1186/1756-9966-29-144

Figure Lengend Snippet: Cycle threshold (Ct) values of candidate reference genes divided in the four tissue groups.

Article Snippet: Transferrin receptor (p90, CD71) , TFRC , Hs99999911_m1 , 105 , Cellular uptake of iron.

Techniques:

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Evaluation of suitable reference genes for normalization of real-time reverse transcription PCR analysis in colon cancer

doi: 10.1186/1756-9966-29-144

Figure Lengend Snippet: Cycle threshold (Ct) values of candidate endogenous control genes across all tissue samples.

Article Snippet: Transferrin receptor (p90, CD71) , TFRC , Hs99999911_m1 , 105 , Cellular uptake of iron.

Techniques: Control, Standard Deviation

Journal: Nature Communications

Article Title: Plasma-derived extracellular vesicles from Plasmodium vivax patients signal spleen fibroblasts via NF-kB facilitating parasite cytoadherence

doi: 10.1038/s41467-020-16337-y

Figure Lengend Snippet: a BCA (protein concentration) of circulating EVs from healthy donors ( h EVs) and P. vivax patients ( Pv EVs) ( n = 10, individual samples). Data show mean ± SD. Two-sided, Mann–Whitney test, * p = 0.0232 (GraphPad). b P. vivax proteins identified by different unique peptides (sequences below the description of corresponding proteins). UniProtKB accession numbers and gene name corresponding to the P. vivax PvP01 strain are shown. c Western blot analysis of Pv EVs obtained from different individual patients (Pv1-7 and Pv10) and human donors (H1 and H3) using anti P. vivax MSP3.1 (upper membrane) and PHISTc (bottom membrane) antibodies. MSP3.1 and PHISTc recombinant truncated-proteins fused to GST, were used as positive controls. Molecular weight in kDaltons (kDa) is shown to the left. M: molecular size marker. Image representative of three independent experiments. d Nanoparticle tracking analysis (NTA) profile (size [nm] versus concentration [particles/mL]) of pooled h EVs and Pv EVs was analysed using NanoSight LM10-12. Table shows mean of three measurement of pooled h EVs and Pv EVs quantified by NTA (particles concentration) and BCA (protein concentration). e Beads-based flow cytometry analysis of pooled h EVs and Pv EVs using six EV markers (CD9, CD63, CD81, GAL3, CD5L and CD71). Data show median fluorescence intensity (MFI) of each antibody and control antibodies (rabbit and mouse-isotype) ± SD (technical replicates, n = 3). Unpaired and two-sided, t -test ** p = 0.0032 (CD63), *** p = 0.0006 (CD81, CD71), **** p < 0.0001 (CD9) (GraphPad). Source data are provided as a Source data file and Supplementary Data , and .

Article Snippet: Further, cells were stained using the following antibodies: CD90-PE/Cy7 [5E10] (Biolegend, Cat#328124) 1/100; CD44-FITC [KM201] (Abcam Cat#ab25340) 1/100; CD54-Alexa Fluor® 488 [HCD54] (Biolegend, Cat#322714) 1/400; CD71-PE [AC102] (Miltenyi Biotec, Cat#130-091-728) 1/400; CD45-PerCP [2D1] (BD Biosciences, Cat#345809) 1/50.

Techniques: Protein Concentration, MANN-WHITNEY, Western Blot, Membrane, Recombinant, Molecular Weight, Marker, Concentration Assay, Flow Cytometry, Fluorescence, Control

Journal: Scientific Reports

Article Title: Tissue-targeted R-spondin mimetics for liver regeneration

doi: 10.1038/s41598-020-70912-3

Figure Lengend Snippet: Design and characterization of cell type specific Wnt signaling enhancing mimetic molecules. ( A ) Scheme of designed molecules. Fragments of RSPO2 spanning the Fu1 and Fu2 domains (action module) were fused the C-terminus of cell type specific or control scFv (targeting module). Specific mutations used are indicated. ( B ) In vitro STF activity of four RSPO proteins. Furin domains of human RSPO1-4 were tested as fusions to an anti-GFP scFv. RSPO2 furin domain alone (N37-E143) was used as a control. Luciferase activity unit is arbitrary (RLU). ( C ) STF activity of RSPO mimetics on HEK293 ( top ) or HuH-7 ( bottom ) in the presence ( left ) or absence ( right ) of an exogenous Wnt source (30% Wnt3a conditioned media). ( D ) Quantitative PCR analysis of ASGR1 , ASGR2 and TFRC gene expression in HEK293, HuH-7 and A431 cells. The signals were relative to ACTB . ( E ) Western Blot analysis on LRP6 receptor and DVL2 phosphorylation in HuH-7 cells. Asterisk indicates a non-specific band that is detected in HuH-7 by the antibody used. The lower bands indicated by the arrow correspond to DVL2 with and without phosphorylation. Tubulin (TUB) is used as the loading control. 10% Wnt3a conditioned media was used as the exogenous Wnt source. Images were cropped for clarity. The uncropped images are presented in Fig. .

Article Snippet: For overexpression of exogenous receptors, cells were transiently transfected with plasmids containing receptors of interest under eukaryotic expression promoters ( ASGR1 was clone OHU03658D from GenScript, ASGR2 was in-house cloned isoform d/NP_001188281.1, and TFRC was clone HG11020-UT from SinoBiologicals), then split into 96-well plates (20,000 cells per well) for STF assay 24 h post transfection.

Techniques: In Vitro, Activity Assay, Luciferase, Real-time Polymerase Chain Reaction, Expressing, Western Blot

Journal: Scientific Reports

Article Title: Tissue-targeted R-spondin mimetics for liver regeneration

doi: 10.1038/s41598-020-70912-3

Figure Lengend Snippet: Confirmation of receptor-specific activation of Wnt signaling. ( A ) STF activity in HEK293 cells transiently transfected with TFR1 (control, top ), ASGR1 ( middle ), and ASGR1 together with ASGR 2 cDNAs ( bottom ), either in the presence ( left ) or absence ( right ) of exogenous Wnt source (30% Wnt3a conditioned media). ( B ) STF activity in A431 cells with TFR1 or ASGR1 overexpression as specified. ( C ) Flow cytometry analysis of cell surface level of FZD proteins. HEK293 cells were transiently transfected with either ZNRF3 alone ( top ) or ASGR1 and ZNRF3 cDNAs ( bottom ), treated with RSPO mimetics as specified at 10 nM, then stained by the pan-FZD antibody 18R5.

Article Snippet: For overexpression of exogenous receptors, cells were transiently transfected with plasmids containing receptors of interest under eukaryotic expression promoters ( ASGR1 was clone OHU03658D from GenScript, ASGR2 was in-house cloned isoform d/NP_001188281.1, and TFRC was clone HG11020-UT from SinoBiologicals), then split into 96-well plates (20,000 cells per well) for STF assay 24 h post transfection.

Techniques: Activation Assay, Activity Assay, Transfection, Over Expression, Flow Cytometry, Staining

Journal: Scientific Reports

Article Title: Tissue-targeted R-spondin mimetics for liver regeneration

doi: 10.1038/s41598-020-70912-3

Figure Lengend Snippet: In vivo tissue-specific enhancement of Wnt signaling. ( A ) STF activity of the RSPO mimetic molecules in the appended-IgG format. ( B ) BLI analysis of the ASGR1 antibody (expressed as a Fab) on human and mouse ASGR1. The affinity (Kd) determined by steady-state fitting is indicated. ( C ) Axin2 and ( D ) Ki67 expression in liver ( upper panel ) and small intestine ( lower panel ) were analyzed by quantitative PCR, 48 h after i.p. injection of proteins as specified (n = 8 mice per group). Statistical analysis was performed using 1-way ANOVA: (ns) not significant, ** p < 0.01, **** p < 0.0001. ( E ) Immunofluorescence staining of liver samples for Ki67 and HNF4α from 10 mg/kg treatment groups. White arrows denote some double-positive cells as examples. DAPI was used to stain nuclei.

Article Snippet: For overexpression of exogenous receptors, cells were transiently transfected with plasmids containing receptors of interest under eukaryotic expression promoters ( ASGR1 was clone OHU03658D from GenScript, ASGR2 was in-house cloned isoform d/NP_001188281.1, and TFRC was clone HG11020-UT from SinoBiologicals), then split into 96-well plates (20,000 cells per well) for STF assay 24 h post transfection.

Techniques: In Vivo, Activity Assay, Expressing, Real-time Polymerase Chain Reaction, Injection, Immunofluorescence, Staining