rabbit monoclonal anti- cdx2  (Cell Signaling Technology Inc)

Journal: Cell Genomics

Article Title: Interpreting regulatory mechanisms of Hippo signaling through a deep learning sequence model

doi: 10.1016/j.xgen.2025.100821

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-CDX2 , Bethyl Laboratories , A300-692A.

Techniques: Immunofluorescence, Recombinant, Ligation, Gel Extraction, Polymerase Chain Reaction, Hybridization, Cloning, Luciferase, CRISPR, Plasmid Preparation, Software, Sonication, Microscopy

Journal: Biology of Reproduction

Article Title: Zygotic activin A is dispensable for the mouse preimplantation embryo development and for the derivation and pluripotency of embryonic stem cells ^†

doi: 10.1093/biolre/ioae156

Figure Lengend Snippet: Embryos deprived of zygotic and exogenous activin A undergo normal development to blastocyst stage. (A, B) Comparison of main morphokinetic parameters between Inhba KO/KO and control— Inhba KO/WT and Inhba WT/WT embryos in C57BL/6/Tar x CBA/Tar (A) and C57BL/6/Tar (B) genetic background. The value of individual parameters was calculated as the number of hours from the injection of hCG to the time of: t NEBD —breakdown of nuclear envelopes of pronuclei; t 2 –t 8 —complete division to the certain number of blastomeres; tM—compaction; tC and tH—beginning of cavitation and hatching process, respectively. On all graphs, the total number of embryos analyzed for each genotype is provided in parentheses. On the boxplots, the middle lines represent medians, the cross shows the mean value, the hinges indicate the interquartile range, the whiskers represent the minimum and maximum values in the group, and the dots show outliers. The lack of statistically significant ( P > 0.05) changes was proved by the Kruskal–Wallis test separately for each morphokinetic parameter. (C) Confocal images of blastocysts in C57BL/6/Tar x CBA/Tar genetic background immunostained with antibodies against CDX2 (TE), SOX17 (PE), Nanog (EPI), and chromatin stained with Chromomycin A3. Scale bars: 20 μm. (D–F) Percentage contribution of primary cell lineages, negative and bipotential cells within blastocyst and inner cell mass in: 120 h C57BL/6/Tar x CBA/Tar blastocysts (D), 135 h C57BL/6/Tar x CBA/Tar blastocysts (E), and 120 h C57BL/6/Tar blastocysts (F) ( P > 0.05, Kruskal–Wallis test). Abbreviations: trophectoderm (TE), inner cell mass (ICM), primitive endoderm (PE), epiblast (EPI), negative (neg.; CDX2−SOX17−Nanog−) cells, bipotential (bipot.; Nanog+SOX17+) cells. See also , and and .

Article Snippet: The following primary antibodies were used: mouse monoclonal antibody (1:50, BioGenex, MU392A-UC; RRID:AB_2923402) or rabbit monoclonal (1:200, Cell Signaling Technology, 12306S; RRID:AB_2797879) antibody against CDX2, rabbit polyclonal antibody against SOX2 (1:100, Abcam, ab97959; RRID:AB_2341193), rat monoclonal antibody against Nanog (1:200, ThermoFisher Scientific, 14-5761-80; RRID:AB_763613), goat polyclonal antibody against SOX17 (1:100, R&D Systems, AF1924; RRID:AB_355060), goat polyclonal antibody against GATA4 (1:100, R&D Systems, AF2606; RRID:AB_2232177), mouse monoclonal antibody against OCT4 (1:100, Santa Cruz Biotechnology, sc-5279; RRID:AB_628051), rat monoclonal antibody against TROMA1 (1:50, Developmental Studies Hybridoma Bank, AB_531826; RRID:AB_531826), goat polyclonal antibody against Brachyury (1:200, R&D Systems, AF2085; RRID:AB_2200235), and rabbit monoclonal antibody against integrin α2 (1:100, Abcam, ab181548; RRID:AB_2847852).

Techniques: Comparison, Control, Injection, Staining

Journal: Cancer Discovery

Article Title: Epigenetic and Oncogenic Inhibitors Cooperatively Drive Differentiation and Kill KRAS - Mutant Colorectal Cancers

doi: 10.1158/2159-8290.CD-23-0866

Figure Lengend Snippet: EZH2 and MEK inhibitors cooperatively suppress WNT signaling and drive differentiation. A, Representative images of vehicle-, MEKi-, EZH2i-, and combo-treated AKP tumor organoids 24 hours after combination treatment (6 days after EZH2i pretreatment). Magnification shown for EZH2i-treated organoids. Scale bars, 150 μm. B, Proliferation assay for AKP and AKPT tumor organoids after treatment with the indicated compounds for 3 days. C, ssGSEA of signatures associated with differentiated cell types using RNA-seq from LOVO and SK-CO1 cell lines treated with EZH2i and/or MEKi. Signatures composed of combined gene lists as described in refs. , . D, Immunoblot of LOVO protein lysates from cells treated with EZH2i and/or MEKi depicting expression of several proteins associated with differentiated intestinal cells (ATOH1, KRT20, KLF4, and CDX2) and stem cells (LGR5, SOX9, PROM1, and CDCA7). pERK and H3K27me3 indicate MEKi and EZH2i target inhibition, and ERK, Histone H3, and GAPDH are loading controls. Quantification indicates fold change (FC) compared with DMSO. E and F, Plots depicting ssGSEA z -scores of signatures associated with oncogenic intestinal Wnt signaling from RNA-seq data in ( E ) SW620 and SK-CO1 cells or ( F ) LOVO cells transduced with an empty vector or a construct to express a constitutively active form of β-catenin after treatment with the indicated agents (C, combination; D, DMSO; E, tazemetostat; M, trametinib). G, Caspase 3/7+ cells (apoptotic) after treatment with EZH2i and/or MEKi in LOVO cells expressing empty vector or β-catenin △90 as measured by Incucyte live-cell imaging. P value determined with two-way ANOVA between empty vector and β-catenin △90 transduced combo-treated cells. H, Proliferation of LOVO pLV β-catenin △90 cells treated with MEKi and/or EZH2i for 5 days. I, Proliferation of LOVO cells transfected with siRNAs against a control sequence or CDX2 and then treated with MEKi and/or EZH2i for 5 days. J, Proliferation of SK-CO1 cells transduced with lentivirus to express LacZ or SOX9 and then treated with MEKi and/or EZH2i for 5 days. Unless otherwise indicated, for all subfigures bars represent mean ± SD of technical replicates. P value measured by unpaired t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: The following antibodies were used at 1:500 to 1:1,000 dilution unless otherwise stated: pERK (CST, #4370, RRID: AB_2315112), ERK (CST, #9102, RRID:AB_330744), H3K27me3 (CST, #9733, RRID:AB_2616029), Histone H3 (CST, #4499, RRID:AB_10544537), TLE4 (Abcam, #ab64833, RRID:AB_2203850), HA-tag (CST, #3724, RRID:AB_1549585), ATOH1 (Proteintech, #21215-1-AP, RRID:AB_10733126), LGR5 (Abcam, #75850, RRID:AB_1523716), SOX9 (EMD Millipore, #AB5535 1:5,000, RRID:AB_2239761), PROM1 (CST, #64326, RRID:AB_2721172), CDX2 (CST, #12306, RRID:AB_2797879), KRT20 (CST, #13063, RRID:AB_2798106), KLF4 (CST, #4038, RRID:AB_2265207), CDCA7 (Proteintech, #15249-1-AP, RRID:AB_2878119), GAPDH (CST, #2118, RRID:AB_561053), vinculin (Santa Cruz Biotechnology, #sc-25336, RRID:AB_628438), β-catenin (CST, #8814, RRID:AB_11127203), p-RB (CST, #9308, RRID:AB_331472), RB (CST, #9313S, RRID:AB_1904119), p21 (CST, #2947, RRID:AB_823586), and p27 (SantaCruz, #sc-528, RRID:AB_632129).

Techniques: Proliferation Assay, RNA Sequencing, Western Blot, Expressing, Inhibition, Transduction, Plasmid Preparation, Construct, Live Cell Imaging, Transfection, Control, Sequencing

Journal: Cancer Discovery

Article Title: Epigenetic and Oncogenic Inhibitors Cooperatively Drive Differentiation and Kill KRAS - Mutant Colorectal Cancers

doi: 10.1158/2159-8290.CD-23-0866

Figure Lengend Snippet: Co-suppression of EZH2 and RAS signaling selectively drives differentiation of tumors in vivo . A, Histological (H&E) and multiplexed immunofluorescent (CyCIF) imaging of cell line–derived xenograft LOVO tumors treated with vehicle or EZH2i for 7 days followed by EZH2i + MEKi for 1, 2, or 3 additional days. Imaging was conducted using antibodies against SMA (cyan; structural), panCK (blue; epithelial), SOX9 (red; stem-like), and CDX2 (green; differentiated) in merged or single channel images as indicated. B, Same as A , but using PDX COCA30 tumors treated with vehicle or EZH2i for 7 days followed by EZH2i + MEKi for 1 additional day. Low- and high-magnification images are shown. C, Multiplexed immunofluorescent imaging (CyCIF) of colon tissue from tumor bearing mice after 21 days of treatment with vehicle or EZH2i + MEKi stained with antibodies for SMA (cyan; structural), Hoechst (blue; nuclei), SOX9 (red; stem-like), CDX2 (green; differentiated), and KRT20 (magenta; differentiated). D, Quantification of Ki67 intensity in cellular compartment of colons described in C . H&E, hematoxylin and eosin.

Article Snippet: The following antibodies were used at 1:500 to 1:1,000 dilution unless otherwise stated: pERK (CST, #4370, RRID: AB_2315112), ERK (CST, #9102, RRID:AB_330744), H3K27me3 (CST, #9733, RRID:AB_2616029), Histone H3 (CST, #4499, RRID:AB_10544537), TLE4 (Abcam, #ab64833, RRID:AB_2203850), HA-tag (CST, #3724, RRID:AB_1549585), ATOH1 (Proteintech, #21215-1-AP, RRID:AB_10733126), LGR5 (Abcam, #75850, RRID:AB_1523716), SOX9 (EMD Millipore, #AB5535 1:5,000, RRID:AB_2239761), PROM1 (CST, #64326, RRID:AB_2721172), CDX2 (CST, #12306, RRID:AB_2797879), KRT20 (CST, #13063, RRID:AB_2798106), KLF4 (CST, #4038, RRID:AB_2265207), CDCA7 (Proteintech, #15249-1-AP, RRID:AB_2878119), GAPDH (CST, #2118, RRID:AB_561053), vinculin (Santa Cruz Biotechnology, #sc-25336, RRID:AB_628438), β-catenin (CST, #8814, RRID:AB_11127203), p-RB (CST, #9308, RRID:AB_331472), RB (CST, #9313S, RRID:AB_1904119), p21 (CST, #2947, RRID:AB_823586), and p27 (SantaCruz, #sc-528, RRID:AB_632129).

Techniques: In Vivo, Imaging, Derivative Assay, Staining