foxo3 antibody  (Bio-Techne corporation)

Journal: Theranostics

Article Title: Microbial metabolite restricts 5-fluorouracil-resistant colonic tumor progression by sensitizing drug transporters via regulation of FOXO3-FOXM1 axis

doi: 10.7150/thno.70754

Figure Lengend Snippet: List of the antibodies and source.

Article Snippet: FOXO3 , Novus Biologicals , NB100-614 , WB , 1:1000.

Techniques:

Journal: bioRxiv

Article Title: Mitochondria define intestinal stem cell differentiation downstream of a FOXO/Notch axis

doi: 10.1101/777391

Figure Lengend Snippet: a. Foxo1 and Foxo3 expression in doxycycline treated shLuc and shFoxo1/3 organoids by qPCR (mean± SEM, n=4) (see also for protein expression). b. Western blot analysis of shluc and shFoxo1/3 organoids, VINCULIN and GAPDH are used as loading controls (representative for n=3). c. Gene expression analysis of shLuc organoids by qPCR and normalized by the non-treated condition (mean± SEM, n=3-5. ANOVA and Dunns multiple comparison test). d. Representative images of doxycycline treated shLuc and shFoxo1/3 organoids cultured in WENR medium (scale bar= 500 AU) and quantification of their perimeter (one representative experiment of 3, each dot represents one organoid, ANOVA and Dunn’s multiple comparison test) e. Flow cytometry gating strategy for quantification of live PCs. f. lmmunohistochemistry of crypts of Villlin-g Cre-;Fox01,3,4L/L and Villin-Cre+ Fox0 1,3,4L/L mice. Proliferative cells are marked by BrdU positive staining.

Article Snippet: Western blot analysis was performed with primary antibodies recognizing: GAPDH (Millipore MAB374), vinculin (Sigma V9131), FOXO1 (Cell Signaling 2880), FOXO3 (Novus Biologicals NB100614), lysozyme (Dako A0099), Tubulin (Millipore CP06 OS), Fis1 (SC376469 Santa Cruz), Cleaved Notch1 (Val1744) (Cell Signaling 4147), OLFM4 (Cell Signaling 39141), Secondary HRP-conjugated antibodies targeting mouse and rabbit IgG were purchased from Biorad.

Techniques: Expressing, Western Blot, Comparison, Cell Culture, Flow Cytometry, Staining

Journal: bioRxiv

Article Title: Mitochondria define intestinal stem cell differentiation downstream of a FOXO/Notch axis

doi: 10.1101/777391

Figure Lengend Snippet: a. t-SNE visualization of 1,522 single cells full-length sc-RNAseq data. Cluster annotation was done on the expression of known cell type markers (See Suppl. Fig. 6a). b. Representation of cell clusters and pseudo-time overlaid on the top 3 Diffusion Components (DC). DC were computed with destiny. Pseudo-time was calculated with Palantir. c. Validation of pseudo-time computation. Gene trends of cell type markers are plotted on pseudo-time for each cell cluster. d. Expression levels of Foxo1, 3 and mitochondrial markers (Hspd1 and Vdac) overlaid on the top 3 DC. Foxo1, 3 and mitochondrial gene trends are plotted on pseudo-time for each cell type cluster as a proxy of gene expression dynamics during cell differentiation. e. 3 dimensional representation of gene expression of Foxo1, Foxo3 and Notch1 in intestinal cells. As a fourth dimension cluster classification is assigned by color (right) or colored according Vdac1 expression levels (left).

Article Snippet: Western blot analysis was performed with primary antibodies recognizing: GAPDH (Millipore MAB374), vinculin (Sigma V9131), FOXO1 (Cell Signaling 2880), FOXO3 (Novus Biologicals NB100614), lysozyme (Dako A0099), Tubulin (Millipore CP06 OS), Fis1 (SC376469 Santa Cruz), Cleaved Notch1 (Val1744) (Cell Signaling 4147), OLFM4 (Cell Signaling 39141), Secondary HRP-conjugated antibodies targeting mouse and rabbit IgG were purchased from Biorad.

Techniques: Expressing, Diffusion-based Assay, Cell Differentiation

Journal: bioRxiv

Article Title: Mitochondria define intestinal stem cell differentiation downstream of a FOXO/Notch axis

doi: 10.1101/777391

Figure Lengend Snippet: a. Visualization of the mean expression of marker genes for each cell lineage plotted on t-SNE well based/ full-length sc-RNAseq data. b. Branch probabilities for each cluster overlaid on the top 1-3 or 1, 2 and 6 DC. Branch probabilities were calculated with Palantir. c. Foxo1, 3 and mitochondrial gene expression levels are plotted on t-SNE plots and boxplots are plotted to represent the quantification of gene expression in each cluster gene. Median values and 25th and 75th percentiles. Mann–Whitney U test, ****= p < 0.0001). d. Validation of sc-RNAseq analysis was done by analyzing a second droplet-based sequencing data of 10396 cells (from Harber et al 2017) t-SNE visualization of droplet-based sequencing of Cluster annotation was done on the expression of known cell type markers (same as in a), and Foxo1, 3 and mitochondrial gene expression levels are plotted on t-SNE and boxplots are plotted to represent the quantification of gene expression in each cluster gene. Median values and 25th and 75th percentiles. Mann–Whitney U test, ****= p < 0.0001). e. 3 dimensional representation of droplet-based sequencing data gene expression of Foxo1, Foxo3 and Notch1 in intestinal cells. As a fourth dimension cluster classification is assigned by color (left) or colored according Vdac1 expression levels (right). f. 3 dimensional representation of well-based scRNA-seq data and second data set of droplet-based scRNA-seq. Genes Ccne1, Cdh1 and Ccnd1 were used as control for comparison with Notch1, Foxo1 and Foxo3. Cluster classification is assigned by color (see c and d).

Article Snippet: Western blot analysis was performed with primary antibodies recognizing: GAPDH (Millipore MAB374), vinculin (Sigma V9131), FOXO1 (Cell Signaling 2880), FOXO3 (Novus Biologicals NB100614), lysozyme (Dako A0099), Tubulin (Millipore CP06 OS), Fis1 (SC376469 Santa Cruz), Cleaved Notch1 (Val1744) (Cell Signaling 4147), OLFM4 (Cell Signaling 39141), Secondary HRP-conjugated antibodies targeting mouse and rabbit IgG were purchased from Biorad.

Techniques: Expressing, Marker, MANN-WHITNEY, Sequencing, Comparison