Journal: The Journal of pediatrics

Article Title: Whole exome sequencing reveals a novel mutation in CUL7 in a patient with an undiagnosed growth disorder

doi: 10.1016/j.jpeds.2012.07.055

Figure Lengend Snippet: Proband’s Diagnostic Genetic Testing

Article Snippet: Whole exome sequencing of the proband, his brother, and both parents was performed at the Broad Institute.

Techniques: Diagnostic Assay, Microarray, Sequencing, Methylation

Journal: International Journal of Chronic Obstructive Pulmonary Disease

Article Title: Identifying targets for COPD treatment through gene expression analyses

doi:

Figure Lengend Snippet: Summary of gene expression profiling studies involving human COPD/emphysema samples

Article Snippet: COPD GOLD2 vs. GOLD0 , Whole lung , Incyte Unigem high-density microarray (10,000 transcripts) SAGE (59,000 tags) , Apoptosis-related genes Inflammation-related genes Transcription factors (increased) Collagens (decreased) , Egr-1/Fos CTGF, CYR61, CX3CL1, TGFB1, and PDGFRA , .

Techniques: Expressing, Functional Assay, Microarray

Journal: Reproductive biology and endocrinology : RB&E

Article Title: Uterine extracellular matrix components are altered during defective decidualization in interleukin-11 receptor α deficient mice

doi: 10.1186/1477-7827-2-76

Figure Lengend Snippet: Real-time RT-PCR amplification conditions for each primer pair

Article Snippet: Primary antibodies used were rabbit anti-mouse collagen type III (Abcam #ab7778, Cambridge, UK) at 5 μg/ml, rabbit anti-mouse biglycan (LF-159 [ ], gift from Dr. Larry Fisher, Matrix Biochemistry Unit, National Institutes of Health, Bethesda, MD) at 1:1000 dilution of whole serum, goat anti-mouse SPARC (Santa Cruz Biotechnology #sc13326, Santa Cruz, CA) at 5 μg/ml, rat anti-mouse entactin/nidogen-1 (Lab Vision-NeoMarkers #RT797P, Fremont, CA) at 15 μg/ml and goat anti-mouse desmin (Santa Cruz) at 200 μg/ml.

Techniques: Quantitative RT-PCR, Amplification

Journal: Reproductive biology and endocrinology : RB&E

Article Title: Uterine extracellular matrix components are altered during defective decidualization in interleukin-11 receptor α deficient mice

doi: 10.1186/1477-7827-2-76

Figure Lengend Snippet: Oligonucleotide primers used in real-time RT-PCR. Primer pairs previously published – COL3A1 [82], BGN [83], SPARC [84] and NID1 [85].

Article Snippet: Primary antibodies used were rabbit anti-mouse collagen type III (Abcam #ab7778, Cambridge, UK) at 5 μg/ml, rabbit anti-mouse biglycan (LF-159 [ ], gift from Dr. Larry Fisher, Matrix Biochemistry Unit, National Institutes of Health, Bethesda, MD) at 1:1000 dilution of whole serum, goat anti-mouse SPARC (Santa Cruz Biotechnology #sc13326, Santa Cruz, CA) at 5 μg/ml, rat anti-mouse entactin/nidogen-1 (Lab Vision-NeoMarkers #RT797P, Fremont, CA) at 15 μg/ml and goat anti-mouse desmin (Santa Cruz) at 200 μg/ml.

Techniques: Quantitative RT-PCR

Journal: Reproductive biology and endocrinology : RB&E

Article Title: Uterine extracellular matrix components are altered during defective decidualization in interleukin-11 receptor α deficient mice

doi: 10.1186/1477-7827-2-76

Figure Lengend Snippet: Gene expression in IL11Ra +/+ and IL11Ra -/- uterus following artificial decidualization. Expression profiling of 15K genes between IL11Ra +/+ and IL11Ra -/- at 0 h (A, B), 18 h (C, D), 24 h (E, F) and 48 h (G, H) following the artificial induction of decidualization. Each volcano style plot shows the normalized log ratio (effect estimate) of IL11Ra -/- compared to wild type for each gene from a series of 4 microarrays, plotted against the log odds of differential expression. A, C, E, G represent the first replicates, and B, D, F, H the second dye-swapped replicates. Genes with log odds of differential expression greater than 3 (ie. adjusted p -value < 0.05, above horizontal line) are represented by open circles, and COL3A1, BGN, SPARC and NID1 are labeled in G and H. Only those genes with log odds of differential expression greater than 3 in both replicates were considered differentially expressed, as described in Methods .

Article Snippet: Primary antibodies used were rabbit anti-mouse collagen type III (Abcam #ab7778, Cambridge, UK) at 5 μg/ml, rabbit anti-mouse biglycan (LF-159 [ ], gift from Dr. Larry Fisher, Matrix Biochemistry Unit, National Institutes of Health, Bethesda, MD) at 1:1000 dilution of whole serum, goat anti-mouse SPARC (Santa Cruz Biotechnology #sc13326, Santa Cruz, CA) at 5 μg/ml, rat anti-mouse entactin/nidogen-1 (Lab Vision-NeoMarkers #RT797P, Fremont, CA) at 15 μg/ml and goat anti-mouse desmin (Santa Cruz) at 200 μg/ml.

Techniques: Gene Expression, Expressing, Quantitative Proteomics, Labeling

Journal: Reproductive biology and endocrinology : RB&E

Article Title: Uterine extracellular matrix components are altered during defective decidualization in interleukin-11 receptor α deficient mice

doi: 10.1186/1477-7827-2-76

Figure Lengend Snippet: Differentially expressed genes in IL11Ra -/- uterus compared to wild type at 48 h of decidualization

Article Snippet: Primary antibodies used were rabbit anti-mouse collagen type III (Abcam #ab7778, Cambridge, UK) at 5 μg/ml, rabbit anti-mouse biglycan (LF-159 [ ], gift from Dr. Larry Fisher, Matrix Biochemistry Unit, National Institutes of Health, Bethesda, MD) at 1:1000 dilution of whole serum, goat anti-mouse SPARC (Santa Cruz Biotechnology #sc13326, Santa Cruz, CA) at 5 μg/ml, rat anti-mouse entactin/nidogen-1 (Lab Vision-NeoMarkers #RT797P, Fremont, CA) at 15 μg/ml and goat anti-mouse desmin (Santa Cruz) at 200 μg/ml.

Techniques: Clinical Proteomics, Sequencing, Reverse Transcription

Journal: Reproductive biology and endocrinology : RB&E

Article Title: Uterine extracellular matrix components are altered during defective decidualization in interleukin-11 receptor α deficient mice

doi: 10.1186/1477-7827-2-76

Figure Lengend Snippet: Quantitative real-time RT-PCR for extracellular matrix components. Quantitative real-time RT-PCR for (A) COL3A1, (B) BGN, (C) SPARC and (D) NID1. Circled data points indicate samples used in the cDNA microarray analysis, and horizontal lines the mean of each genotype. Absolute values for mRNA abundance were normalized to that of 18S rRNA.

Article Snippet: Primary antibodies used were rabbit anti-mouse collagen type III (Abcam #ab7778, Cambridge, UK) at 5 μg/ml, rabbit anti-mouse biglycan (LF-159 [ ], gift from Dr. Larry Fisher, Matrix Biochemistry Unit, National Institutes of Health, Bethesda, MD) at 1:1000 dilution of whole serum, goat anti-mouse SPARC (Santa Cruz Biotechnology #sc13326, Santa Cruz, CA) at 5 μg/ml, rat anti-mouse entactin/nidogen-1 (Lab Vision-NeoMarkers #RT797P, Fremont, CA) at 15 μg/ml and goat anti-mouse desmin (Santa Cruz) at 200 μg/ml.

Techniques: Quantitative RT-PCR, Microarray

Journal: Reproductive biology and endocrinology : RB&E

Article Title: Uterine extracellular matrix components are altered during defective decidualization in interleukin-11 receptor α deficient mice

doi: 10.1186/1477-7827-2-76

Figure Lengend Snippet: Immunohistochemistry for extracellular matrix components. Immunohistochemical staining of wild type (A, C, E, G, I, K, M, O, P, Q, S) and IL11Ra -/- (B, D, F, H, J, L, N, R, T) uterus at 48 h of decidualization using specific antibodies for collagen III (A, B, C, D), biglycan (E, F, G, H), nidogen-1 (I, J, K, L), SPARC (M, N, O, P) and desmin (Q, R, S, T). Negative controls using a matching concentration of non-immune IgG (collagen III, nidogen-1, SPARC and desmin) or normal serum (biglycan) in place of the primary antibody are inset in A, B, E, F, I, J, M, N, Q and R. Black squares on A and B indicate the antimesometrial pole magnified in C and D. Abbreviations: connective tissue (ct), myometrium (my), mesometrial pole (m), antimesometrial pole (am), luminal epithelium (le), glandular epithelium (ge), decidualized stromal cell (dsc), non-decidualized stromal cell (sc), blood vessel (bv), glycocalyx (gly). Scale bar = 50 μm (A, B, E, F, I, J, M, N, Q and R are at the same magnification; C, D, G and P are at the same magnification; H, K, L, O, S, T and inset in G are at the same magnification).

Article Snippet: Primary antibodies used were rabbit anti-mouse collagen type III (Abcam #ab7778, Cambridge, UK) at 5 μg/ml, rabbit anti-mouse biglycan (LF-159 [ ], gift from Dr. Larry Fisher, Matrix Biochemistry Unit, National Institutes of Health, Bethesda, MD) at 1:1000 dilution of whole serum, goat anti-mouse SPARC (Santa Cruz Biotechnology #sc13326, Santa Cruz, CA) at 5 μg/ml, rat anti-mouse entactin/nidogen-1 (Lab Vision-NeoMarkers #RT797P, Fremont, CA) at 15 μg/ml and goat anti-mouse desmin (Santa Cruz) at 200 μg/ml.

Techniques: Immunohistochemistry, Immunohistochemical staining, Staining, Concentration Assay

Journal:

Article Title: Brain Lipid Binding Protein in Axon-Schwann Cell Interactions and Peripheral Nerve Tumorigenesis

doi: 10.1128/MCB.23.6.2213-2224.2003

Figure Lengend Snippet: mRNA expression analysis in Nf1 mutant mouse Schwann cells. (A) Microarray analysis was used to compare genome-wide expression levels between normal mouse Schwann cells and Nf1 mutant Schwann cells. The control for each comparison was Cy3-labeled cDNA generated from normal mouse Schwann cell mRNA. For each of four Nf1 mutant Schwann cell samples (Nf1+/−, Nf1−/−, Nf1−/− TXF, and Nf1−/− TXF treated with FTI), mRNA was used as a template to synthesize Cy5-labeled cDNA. Cy3- and Cy5-labeled cDNA probes were hybridized simultaneously to the Incyte Genomics MouseGEM 1.0 cDNA microarray. Relative intensities of Cy3 versus Cy5 fluorescent signals for each cDNA target sequence were analyzed with GeneSpring software. The most changes were observed in the Nf1−/− TXF cells (genes upregulated in Nf1−/− TXF are red; genes downregulated in Nf1−/− TXF are green). Expression of one target cDNA, BLBP (black line), was 26-fold above normal in the Nf1−/− TXF cells and not normalized by FTI treatment. (B) RT-PCR analysis confirmedthe microarray result of elevated BLBP expression in Nf1−/− TXF cells. Reverse transcriptase (RT) was omitted from duplicate samples to control for DNA contamination. Primers for BLBP (∼200-bp amplicon) and actin control primers (∼500-bp amplicon) were included in the mixture for each 40-cycle reaction. The plasmid positive control for BLBP amplification is the UniGEM clone (Incyte Genomics) containing the BLBP cDNA insert spotted on the microarray. (C) Quantitative real-time PCR of BLBP normalized to GAPDH resulted in a 145-fold change over expression in Nf1−/− TXF cells compared to wild-type mouse Schwann cells. Rn, fluorescent signal intensity; horizontal starred line, chosen threshold at geometric phase of amplification.

Article Snippet: Expression of exogenous protein was detected by immunolabeling with mouse anti-HA (Santa Cruz) followed by donkey anti-mouse Cy3-conjugated secondary antibodies (Jackson Immunoresearch) or goat anti-human EGFR antibodies (Santa Cruz) followed by donkey anti-goat tetramethyl rhodamine isocyanate-labeled secondary antibodies (Jackson Immunoresearch).

Techniques: Expressing, Mutagenesis, Microarray, Genome Wide, Labeling, Generated, Sequencing, Software, Reverse Transcription Polymerase Chain Reaction, Amplification, Plasmid Preparation, Positive Control, Real-time Polymerase Chain Reaction, Over Expression

Journal:

Article Title: Brain Lipid Binding Protein in Axon-Schwann Cell Interactions and Peripheral Nerve Tumorigenesis

doi: 10.1128/MCB.23.6.2213-2224.2003

Figure Lengend Snippet: Mouse neuron-Schwann cell coculture. Anti-BLBP promotes extension of Nf1−/− TXF cell processes along axons. Wild-type (A and B) or Nf1−/− TXF (C and D) mouse Schwann cells labeled with Cell Tracker green were preincubated with rabbit IgG (A and C) or anti-BLBP antibodies (B and D) and seeded onto DRGN cultures stripped of endogenous Schwann cells. Two days after seeding, cocultures were fixed and stained with antineurofilament antibodies followed by Cy3 (red)-conjugated secondary antibodies. Confocal images obtained with Zeiss LSM Image Browser software are shown. Single cells are representative of the majority observed with each treatment. Arrowheads indicate Schwann cell processes. The asterisk indicates the region which is magnified fivefold in the inset. The scale bar in panel C equals 5 μm and also applies to panels A, B, and D. (E) Lower magnification (scale bar, 5 μm) of Nf1−/− TXF on DRGN cultures in the presence of anti-BLBP antibodies. Arrowheads indicate processes from two cells extending along neurites; other cells lack processes. (F) Extension of Nf1−/− TXF cell processes in the presence of anti-BLBP antibodies is statistically significant. The percentages of Nf1−/− TXF cells extending processes along axons in the presence of control IgG (gray bar) or anti-BLBP antibodies (black bar) are graphed. Error bars reflect standard deviations in a Student t test (P = 0.003).

Article Snippet: Expression of exogenous protein was detected by immunolabeling with mouse anti-HA (Santa Cruz) followed by donkey anti-mouse Cy3-conjugated secondary antibodies (Jackson Immunoresearch) or goat anti-human EGFR antibodies (Santa Cruz) followed by donkey anti-goat tetramethyl rhodamine isocyanate-labeled secondary antibodies (Jackson Immunoresearch).

Techniques: Labeling, Staining, Software

Journal: The Journal of Biological Chemistry

Article Title: Essential Role of the Zinc Finger Transcription Factor Casz1 for Mammalian Cardiac Morphogenesis and Development *

doi: 10.1074/jbc.M114.570416

Figure Lengend Snippet: Casz1 expression pattern during mouse embryogenesis. A, cartoon of the Casz1 gene trap that inserted with a βgeo reporter after Casz1 exon 9 resulting in a truncated Casz1. B, genotyping of Casz1-trapped mice by RT-PCR. C, whole mount X-gal staining (blue) of E9.5 Casz1+/βgeo embryos shows that Casz1 was expressed in the hindbrain, neural tube, and heart, which are further demonstrated by dissected embryo sagittal sections 1, 2, and 3 (100× magnification). Sagittal section 3b (200× magnification) shows that Casz1 was expressed in the cardiomyocytes. D and E, whole mount X-gal staining of E12.5 Casz1+/+, Casz1+/βgeo, and Casz1βgeo/βgeo embryos showed that Casz1 is expressed in the eye, dorsomedial telencephalon, cranial ganglia, nasal placode, somite, neural tube, and heart. F, real time PCR to detect wild type Casz1 allele and Gata4 mRNA levels in E12.5 hearts. G, the protein levels of Casz1a/Casz1b and GAPDH were visualized by immunoblotting E14.5 whole heart lysate with anti-Casz1 antibody and anti-GAPDH antibody. H, whole mount anti-β-galactosidase staining (red) of E14.5 Casz1+/+, Casz1+/βgeo, and Casz1βgeo/βgeo hearts.

Article Snippet: Rabbit anti-GAPDH (Santa Cruz, 1:4000) and rabbit anti-Casz1 (Rockland, 1:1000) were used to detect GAPDH and Casz1 protein level in heart.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Staining, Real-time Polymerase Chain Reaction, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Essential Role of the Zinc Finger Transcription Factor Casz1 for Mammalian Cardiac Morphogenesis and Development *

doi: 10.1074/jbc.M114.570416

Figure Lengend Snippet: Hypoplasia in the myocardium of Casz1βgeo/βgeo heart. A, phenotype of Casz1+/+, Casz1+/βgeo and Casz1βgeo/βgeo E 15.5 embryos. The edema was seen at the neck region on the back of Casz1βgeo/βgeo embryos but not other embryos (green arrow). B–D, column 1, H&E staining of transverse sections of E15.5 Casz1+/+ (B1), Casz1+/βgeo (C1), and Casz1βgeo/βgeo (D1) embryos. Column 2, right ventricle (RV) magnification of the respective green squares from column 1. Column 3, septum (SP) magnification of respective yellow squares from column 1. Column 4, left ventricle (LV) magnification of respective blue squares from column 1. E, representative photomicrographs indicate accumulation of blood cells in the liver of the Casz1βgeo/βgeo embryo (right panel) but not in the Casz1+/+ (left panel) or Casz1+/βgeo (middle panel) embryo.

Article Snippet: Rabbit anti-GAPDH (Santa Cruz, 1:4000) and rabbit anti-Casz1 (Rockland, 1:1000) were used to detect GAPDH and Casz1 protein level in heart.

Techniques: Staining

Journal: The Journal of Biological Chemistry

Article Title: Essential Role of the Zinc Finger Transcription Factor Casz1 for Mammalian Cardiac Morphogenesis and Development *

doi: 10.1074/jbc.M114.570416

Figure Lengend Snippet: Decreased proliferation in Casz1βgeo/βgeo heart. A, the E14.5 Casz1βgeo/βgeo hearts were morphologically different compared with the Casz1+/βgeo or wild type heart (compare the green arrow targeted region). B, lower magnification of H&E staining of transverse sections from E14.5 embryos (upper panels). The lower panels represent a magnification of the regions that are depicted in the blue squares in the upper panel. The green arrow denotes the septal defect apparent in the serial sectioning of the Casz1βgeo/βgeo heart compared with the Casz1+/+ heart. C, paraffin sections from E14.5 hearts from Casz1+/+ and Casz1βgeo/βgeo hearts were immunostained with antibodies for MF20 (red) and the mitosis marker phosphohistone H3 (pH3, green). The nuclei were stained with DAPI (blue; left top panels, original overview; left bottom panels, high magnification). In the high magnification images, arrows indicate pH3+ cells. The graph (right panel) represents the relative percentage of pH3 positive cells compared with the total number of DAPI positive Casz1βgeo/βgeo or Casz1+/+ cardiomyocytes from at least five sections from each embryo. The Casz1βgeo/βgeo cardiomyocytes proliferate significantly slower than Casz1+/+ cardiomyocytes (p < 0.00001). D, tunnel staining (green) was performed using E14.5 heart paraffin sections (upper panels), and nuclei were stained with DAPI (blue). The lower panels represent magnified areas delineated in the red squares in the upper panels. There was no significant difference in the number of apoptotic cells observed in Casz1βgeo/βgeo cardiomyocytes compared with Casz1+/+ cardiomyocytes.

Article Snippet: Rabbit anti-GAPDH (Santa Cruz, 1:4000) and rabbit anti-Casz1 (Rockland, 1:1000) were used to detect GAPDH and Casz1 protein level in heart.

Techniques: Staining, Marker

Journal: The Journal of Biological Chemistry

Article Title: Essential Role of the Zinc Finger Transcription Factor Casz1 for Mammalian Cardiac Morphogenesis and Development *

doi: 10.1074/jbc.M114.570416

Figure Lengend Snippet: Disorganized fiber orientation in Casz1βgeo/βgeo heart. To visualize thin filaments, Casz1+/+ and Casz1βgeo/βgeo E14.5 hearts were stained with phalloidin-488 (green) to detect F-Actin and co-stained with TOPRO3 to show cell nuclei. The left panels show the co-staining results of the left ventricles of the hearts. The right panels represent magnified areas delineated in the red squares in the left panels. Enlarged images showed complete disruption of the fiber orientation/cell alignment in the left ventricle (LV) of Casz1βgeo/βgeo heart.

Article Snippet: Rabbit anti-GAPDH (Santa Cruz, 1:4000) and rabbit anti-Casz1 (Rockland, 1:1000) were used to detect GAPDH and Casz1 protein level in heart.

Techniques: Staining

Journal: The Journal of Biological Chemistry

Article Title: Essential Role of the Zinc Finger Transcription Factor Casz1 for Mammalian Cardiac Morphogenesis and Development *

doi: 10.1074/jbc.M114.570416

Figure Lengend Snippet: Abnormal gene expression in Casz1βgeo/βgeo heart. A, microarray analysis of RNA from three E12.5 Casz1+/+ hearts and three E12.5 Casz1βgeo/βgeo hearts. The heat map was generated using Partek software. Two-thirds of these genes are aberrantly up-regulated, and one-third are aberrantly down regulated in Casz1βgeo/βgeo hearts. B, Partek gene ontology oncology analysis showed that the top two categories of enriched genes are “signaling” and the process of biological adhesion. Biological adhesion genes in Casz1βgeo/βgeo hearts that are up-regulated (red) or down-regulated (blue) are listed below. C, IPA assays showed gene enrichment in the categories of physiological system development and function (panel 1) with a key subcategory being skeletal and muscular system development and function genes that are up-regulated (red) or down-regulated (blue) in Casz1βgeo/βgeo hearts listed below and molecular and cellular function (panel 2) with a key subcategory of genes in molecular transport and the ion transport genes that are up-regulated (red) or down-regulated (blue) listed below. D, verification of microarray results. The mRNA levels of representative genes encoding cell adhesion molecules, muscle contraction, and muscular development proteins and ion channels were evaluated by real time PCR and normalized to GAPDH gene. The mRNA levels of these genes in Casz1βgeo/βgeo hearts were significantly different compared with their levels in Casz1+/+ hearts. The bar graph represents means ± S.D. (all p < 0.005).

Article Snippet: Rabbit anti-GAPDH (Santa Cruz, 1:4000) and rabbit anti-Casz1 (Rockland, 1:1000) were used to detect GAPDH and Casz1 protein level in heart.

Techniques: Expressing, Microarray, Generated, Software, Cell Function Assay, Real-time Polymerase Chain Reaction

Journal: The Journal of Biological Chemistry

Article Title: Essential Role of the Zinc Finger Transcription Factor Casz1 for Mammalian Cardiac Morphogenesis and Development *

doi: 10.1074/jbc.M114.570416

Figure Lengend Snippet: Abnormal expression of genes involved in cell cycle and regulation of cell proliferation in Casz1βgeo/βgeo heart. A, IPA assays showed gene enrichment in the categories of cell cycle and cellular growth and proliferation. The genes that are up-regulated (red) or down-regulated (blue) in Casz1βgeo/βgeo hearts were listed below. B, verification of microarray results. The mRNA levels of representative genes were evaluated by real time PCR and normalized to GAPDH gene. The mRNA levels of these genes in Casz1βgeo/βgeo hearts were significantly different compared with their levels in Casz1+/+ hearts. The bar graph represents means ± S.D. (all p < 0.05).

Article Snippet: Rabbit anti-GAPDH (Santa Cruz, 1:4000) and rabbit anti-Casz1 (Rockland, 1:1000) were used to detect GAPDH and Casz1 protein level in heart.

Techniques: Expressing, Microarray, Real-time Polymerase Chain Reaction

Journal: The Journal of Biological Chemistry

Article Title: Essential Role of the Zinc Finger Transcription Factor Casz1 for Mammalian Cardiac Morphogenesis and Development *

doi: 10.1074/jbc.M114.570416

Figure Lengend Snippet: Validation of Casz1 target genes in cellular models. A, human cardiac fibroblasts were transfected with empty vector (EMV) or CASZ1b vector and cultured for 5 days. Panel a, relative CASZ1b mRNA level was evaluated by real time PCR using human CASZ1 primer. The bar graph represents means ± S.E.(#, p < 0.05). Panel b, Western blot to show the overexpression of Casz1b in CASZ1b overexpressed cells. Panel c, after overexpression of CASZ1b, relative mRNA levels of representative genes were evaluated by real time PCR. The bar graph represents means ± S.E. (#, p < 0.05). B, knockdown or overexpression of Casz1 in HL-1 cells. Panel a, HL-1 cells were transfected with nontargeting control (con) siRNA or Casz1 siRNA, and after a 2-day culture, the relative mRNA levels of representative genes were evaluated by real time. The bar graph represents means ± S.D. (*, p < 0.005). Panel b, HL-1 cells were transfected with EMV or CASZ1b vector, and after a 2-day culture, the relative CASZ1b mRNA levels were evaluated by real time PCR using human CASZ1 primers. The graph represents mean ± S.D. (*, p < 0.005). Panel c, after transfection with CASZ1b and a two-dimensional culture, the relative mRNA levels of representative genes were evaluated by real time PCR. The bar graph represents means ± S.D. (#, p < 0.05; ns represents no statistic significant difference).

Article Snippet: Rabbit anti-GAPDH (Santa Cruz, 1:4000) and rabbit anti-Casz1 (Rockland, 1:1000) were used to detect GAPDH and Casz1 protein level in heart.

Techniques: Transfection, Plasmid Preparation, Cell Culture, Real-time Polymerase Chain Reaction, Western Blot, Over Expression

Journal: The Journal of Biological Chemistry

Article Title: Essential Role of the Zinc Finger Transcription Factor Casz1 for Mammalian Cardiac Morphogenesis and Development *

doi: 10.1074/jbc.M114.570416

Figure Lengend Snippet: Sarcomeric organization in Casz1βgeo/βgeo heart. A, GSEA assay indicated the negative enrichment of genes encoding contractile fiber proteins (panels a and b), as well as genes encoding contractile fiber part proteins (panel c). NES, normalized enrichment score; Nom, nominal; FDR, false discovery rate. B, sections containing the trabecular regions of E14.5 hearts were labeled for desmin (red), and nuclei were stained with DAPI (blue). There is a striated pattern for desmin labeling in both wild type and Casz1-deficient hearts, but the striated patterns are not as uniform or apparent in the Casz1-deficient heart.

Article Snippet: Rabbit anti-GAPDH (Santa Cruz, 1:4000) and rabbit anti-Casz1 (Rockland, 1:1000) were used to detect GAPDH and Casz1 protein level in heart.

Techniques: Labeling, Staining

Journal: The Journal of Biological Chemistry

Article Title: Essential Role of the Zinc Finger Transcription Factor Casz1 for Mammalian Cardiac Morphogenesis and Development *

doi: 10.1074/jbc.M114.570416

Figure Lengend Snippet: Abnormal Z line organization in E14.5 Casz1βgeo/βgeo heart. A, cardiomyocytes isolated from Casz1+/+ and Casz1βgeo/βgeo E14.5 embryo hearts were cultured in vitro for 2 days and immunostained for α-actinin (red) and F-actin (green), whereas the nuclei were stained with DAPI (blue). Representative images show that clear Z lines were presented in the wild type cardiomyocytes, but not in the Casz1βgeo/βgeo cardiomyocytes. B, the CHITEST histogram shows the percentage of Casz1+/+ or Casz1βgeo/βgeo cardiomyocytes with clear Z lines (Z line+) or abnormal Z lines (Z line−) (p < 0.00001). The data are from two independent experiments (>100 Casz1+/+ or Casz1βgeo/βgeo cardiomyocytes were counted in each experiment). C, more representative images show that clear Z lines are present in the Casz1+/+ cardiomyocytes. There are no clear Z lines in these Casz1βgeo/βgeo cardiomyocytes, and the α-actinin staining pattern is distinct compared with the Casz1+/+ cardiomyocytes.

Article Snippet: Rabbit anti-GAPDH (Santa Cruz, 1:4000) and rabbit anti-Casz1 (Rockland, 1:1000) were used to detect GAPDH and Casz1 protein level in heart.

Techniques: Isolation, Cell Culture, In Vitro, Staining

Journal: Cell Reports Medicine

Article Title: CMG901, a Claudin18.2-specific antibody-drug conjugate, for the treatment of solid tumors

doi: 10.1016/j.xcrm.2024.101710

Figure Lengend Snippet:

Article Snippet: ChromPure Mouse IgG,whole molecule , Jackson ImmunoResearch , 015-000-003.

Techniques: Clone Assay, Clinical Proteomics, Microarray, Recombinant, Transfection, Membrane, Integrity Assay, Antibody Labeling, Reverse Transcription, Plasmid Preparation, Software