Journal: bioRxiv

Article Title: Overexpression of CD47 is associated with brain overgrowth in 16p11.2 deletion syndrome

doi: 10.1101/808022

Figure Lengend Snippet: A. Representative histograms show the mean fluorescence intensities (MFI) of cell-surface expression of Calreticulin (CRT) in NPCs derived from control, 16p_del, and 16p_dup lines. B. Quantification of CRT MFI of cell-surface expression of CRT in the 16p_del NPCs relative to 16p_dup and control NPCs. C. Quantification of Phaseolus Vulgaris Leucoagglutinin (PHA-L), indicative of asialoglycan binding sites for CRT, in NPCs in 16p_deletion lines relative to control and 16p_duplication lines. D. Representative histograms show the MFI of cell-surface expression of CRT in OPCs derived from control, 16p_del, and 16p_dup lines. E. Quantification of CRT MFI of cell-surface expression of CRT in the 16p_del OPCs relative to 16p_dup and control OPCs. F. Quantification of PHA-L in OPCs in 16p_deletion lines relative to control and 16p_duplication lines. n=2 biological replicates per cell line (16p_dup, n=2; control, n=3; 16p_del, n=4 cell lines). All data are mean ± SE; P values were determined by one-way ANOVA followed by post-hoc Tukey HSD Test.

Article Snippet: The lectin fluorescein labeled Phaseolus Vulgaris Leucoagglutinin (PHA-L) (1:200; Vector Laboratories) was used to stain for PHA-L. For O4 staining, the cells were first incubated in primary antibody mouse-anti-O4 antibody (Clone 81; 1:100; EMD millipore), followed by secondary antibody incubation, anti-Mouse IgM Alexa Fluor 488 (1:1000; Life Technologies).

Techniques: Fluorescence, Expressing, Derivative Assay, Binding Assay

Journal: Stem cells (Dayton, Ohio)

Article Title: Mesenchymal stem cells and endothelial progenitor cells decrease renal injury in experimental swine renal artery stenosis through different mechanisms

doi: 10.1002/stem.1263

Figure Lengend Snippet: Top: Representative images of CM-DiI labeled (red) endothelial progenitor cells (EPC) or mesenchymal stem cells (MSC) in the post-stenotic kidneys of pigs with renal artery stenosis (RAS) 4 weeks after cell delivery. Green shows peanut agglutinin (PA, green arrow), a distal tubular marker, and cyan shows a proximal tubular marker phaseolus vulgaris erythroagglutinin (PHA-E, cyan arrow). EPC showed mainly tubular engraftment (yellow arrow), while MSC tend to integrate into both proximal tubules (yellow arrow) and interstitial area (red arrow). Bottom: Both EPC and MSC improved RBF and GFR in pigs with RAS, yet MSC more effectively restored GFR. *p<0.05 vs. Normal, †p<0.05 vs. RAS. Scale bar=200µm

Article Snippet: Furthermore, frozen kidney sections from pigs infused with cells were stained with the distal tubular marker peanut agglutinin (5ug/ml, Vector) and proximal tubular marker phaseolus vulgaris erythroagglutinin (PHA-E, 5ug/ml, Vector).

Techniques: Labeling, Marker

Journal: The Journal of Biological Chemistry

Article Title: MicroRNA-424 Predicts a Role for β-1,4 Branched Glycosylation in Cell Cycle Progression

doi: 10.1074/jbc.M115.672220

Figure Lengend Snippet: Loss of MGAT4A activity does not affect GlcNAc-β-1,6 branching. A, biosynthetic pathway of tetraantennary branched N-glycans. Binding sites of PHA-L are indicated. B, analysis of PHA-L binding to the Consortium of Functional Glycomics (CFG) glycan array v. 5.0 indicates a preference for GlcNAc-β-1,6 branched products. The graph shows the data for 100 μg/ml biotinylated-PHA-L (Vector Laboratories) binding to glycans bearing only β-1,4 branching (β-(1,4)) and the corresponding β-1,6 glycans (β-(1,6)). Data are given as fluorescence intensity. C, knockdown of MGAT4A in MCF-7 cells shows no significant change in MGAT4B or MGAT5 mRNA levels by qRT-PCR. The graph shows -fold change mRNA expression compared with NTC. n = 3 biological replicates. Error bars represent the S.D. D, fluorescence microscopy of PHA-L stained MGAT4A-KD and NTC cells. No significant loss of PHA-L binding were observed. Images shown are representative of two biological replicates, five images per replicate.

Article Snippet: The graph shows the data for 100 μg/ml biotinylated-PHA-L (Vector Laboratories) binding to glycans bearing only β-1,4 branching (β -(1,4) ) and the corresponding β-1,6 glycans (β -(1,6) ).

Techniques: Activity Assay, Binding Assay, Functional Assay, Plasmid Preparation, Fluorescence, Quantitative RT-PCR, Expressing, Microscopy, Staining

Journal: bioRxiv

Article Title: Evolution of mechanisms controlling epithelial morphogenesis across animals: new insights from dissociation - reaggregation experiments in the sponge Oscarella lobularis

doi: 10.1101/2021.03.22.436370

Figure Lengend Snippet: (A- C’’) Control conditions in natural sea water (1h NSW): (A) Confocal microscopic view (LM) of a choanocyte chamber, the classical round shape of the choanocyte chamber is visible (dotted yellow line) with its typical collar of apical microvilli (blue arrow). The type IV Collagen staining (in magenta) is lining the basal pole of choanocyte chambers (pink arrow). (A) LM focused view on the actin-rich cellular junction between choanocytes (yellow arrow). (B) Scanning Electron Microscopic (SEM) view of a choanocyte chamber with collar of apical microvilli (cyan arrow), and a basement membrane lining the choanocyte chamber (pink arrow). (C’) Focus on the cell-cell junction (yellow arrow), basal lamina (pink arrow) and apical microvilli (cyan arrow). (C’’) Choanocyte with its complete apical microvilli collar (cyan arrow) and complete basal lamina (pink arrow). (D-F’’’) Choanocyte chambers observed one hour after incubation in calcium-magnesium-free sea water (1hD CMSFW): (D) The general architecture of the choanocyte chamber is still observable in LM (dotted yellow line), immunostaining of type IV Collagen is not visible anymore (dotted pink arrow), apical microvilli are disintegrated (dotted cyan arrow). (E) Focus (LM) on a choanocyte with its disintegrated apical microvilli (dotted cyan arrow) and numerous basal actin protrusions (red arrow). (F) SEM view of a choanocyte chamber. Choanocytes present vacuoles (black arrow), disintegrated apical microvilli (dotted cyan arrow) and protrusions (red arrow). The basal lamina is detached from the choanocytes (dotted pink arrow). (F’) Focus (SEM) on the disintegration of the apical microvilli (cyan dotted arrow) and on the loss of cell-cell contact (dotted yellow arrow). (F’’) Focus on basal actin protrusions at the basal pole of a choanocyte (SEM) (red arrow). (F’’’) Focus on two choanocytes with a clear detachment from the basal lamina (dotted pink arrow). (G-I’’) Choanocyte chambers observed four hours after incubation in calcium-magnesium-free sea water (4hD CMFSW): (G) LM view of a destructured choanocyte chamber, immunostaining of type IV Collagen is not visible and the general architecture of the choanocyte chamber is no more recognizable (yellow dotted line). (H) (LM) Focus on a choanocyte with numerous actin basal protrusions (red arrow). (I ) SEM view of choanocyte showing actin protrusions (red arrow) and the whole destructuring of the basal lamina (dotted pink arrow). (I’) (SEM) focus on the actin protrusions (red arrow). (I’’) The basal lamina is completely lost at this time-point (SEM) (dotted pink arrow). For all confocal (LM) pictures: Staining in grey: Phalloidin, Green: PhaE lectin, Blue: DAPI, Magenta: type IV Collagen. Additional picture of control condition in NSW is available in the Supplementary Figure S2B.

Article Snippet: To monitor choanocytes, fluorescein labeled lectin PhaE ( Phaseolus vulgaris Erythroagglutinin ) (Vector lab) was used to specifically stain this cell type (1: 500; ).

Techniques: Staining, Incubation, Immunostaining