Journal: Cancers

Article Title: Mutant KRAS and GATA6 Stratify Survival in Patients Treated with Chemotherapy for Pancreatic Adenocarcinoma: A Prospective Cohort Study.

doi: 10.3390/cancers17050896

Figure Lengend Snippet: Figure 3. Survival analysis according to the tissue biomarkers. (A) Progression-free survival and (B) overall survival according to GATA6 RNA expression. (C) Progression-free survival and (D) overall survival according to GATA6 tissue expression (IHC). CI: confidence interval; GATA6: GATA binding protein 6; IHC: immunohistochemistry; NA: not assessed.

Article Snippet: The primary antibodies used were rabbit polyclonal antibodies against human DCK (LS-B1825, LifeSpan Bioscience, Inc., Seattle, WA, USA), human hENT1 (11337-1-AP, ProteinTech Group, Inc., Chicago, IL, USA), human CES2 (HPA018897, Atlas Antibodies Inc., Stockholm, Sweden), and anti-GATA6 polyclonal antibody (HPA066629, Atlas Antibodies Inc.) Representative images of the tissue biomarkers are shown in Supplementary Figure S1.

Techniques: RNA Expression, Expressing, Binding Assay, Immunohistochemistry

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Transcription factor GATA6: a novel marker and putative inducer of ductal metaplasia in biliary atresia

doi: 10.1152/ajpgi.00362.2017

Figure Lengend Snippet: Quantitative real-time PCR primer sequences

Article Snippet: Primary antibodies used were as follows: rabbit antihuman GATA6 IgG (sc-9055; Santa Cruz Biotechnologies, Santa Cruz, CA) at dilution 1:1,200 for human samples, goat anti-human GATA6 IgG (AF1700; R&D Systems, Minneapolis, MN) at dilution 1:1,200 for mouse samples.

Techniques: Real-time Polymerase Chain Reaction, Sequencing

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Transcription factor GATA6: a novel marker and putative inducer of ductal metaplasia in biliary atresia

doi: 10.1152/ajpgi.00362.2017

Figure Lengend Snippet: GATA6 is expressed in human hepatocytes during early gestation but not in normal perinatal or adult hepatocytes. A–C: immunohistochemical staining of GATA6 protein in normal human liver. Brown indicates positive staining in nuclei. In fetal liver (FL) at gestational week (GW) 13 (FL GW13), GATA6 protein is expressed in all hepatocytes (arrowhead) and other cell types of the liver (A). At GW37, GATA6 protein expression is diminished in hepatocytes (arrowhead) and restricted to cholangiocytes lining the bile ducts (arrow) (B). The expression pattern of GATA6 protein in adult liver (AL) is similar to that of late FL with positive cholangiocytes (arrow) and negative hepatocytes (arrowhead) (C). D–G: in situ hybridization of GATA6 mRNA in FL GW37 (D and E) and AL (F and G). Green indicates positive signal in cytoplasm. Cholangiocytes show positive signal (arrow); hepatocytes show weak or negative signal (arrowhead). Scale bar = 200 µm (A–C) and 20 µm (D–G).

Article Snippet: Primary antibodies used were as follows: rabbit antihuman GATA6 IgG (sc-9055; Santa Cruz Biotechnologies, Santa Cruz, CA) at dilution 1:1,200 for human samples, goat anti-human GATA6 IgG (AF1700; R&D Systems, Minneapolis, MN) at dilution 1:1,200 for mouse samples.

Techniques: Immunohistochemical staining, Staining, Expressing, In Situ Hybridization

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Transcription factor GATA6: a novel marker and putative inducer of ductal metaplasia in biliary atresia

doi: 10.1152/ajpgi.00362.2017

Figure Lengend Snippet: Hepatocyte expression of GATA6 is elevated in biliary atresia (BA) and decreased after portoenterostomy (PE). A: boxplot showing the relative GATA6 mRNA expression, as determined by qPCR, in different liver sample groups. Dots represent individual samples, the box represents the interquartile range, and the whiskers represent the 1st and 4th quartile. The line inside the box is the median, and the dashed line represents the mean. *P < 0.05, **P < 0.01 compared with BA group. The P values of other paired comparisons are indicated in Table 2. B–E: in situ hybridization of GATA6 in BA samples demonstrates strongly positive hepatocytes (arrowhead) (B and C), whereas in situ hybridization in BA-post-PE samples shows weak or negligible signal in hepatocytes (arrowhead) (D and E). F–H: GATA6 immunohistochemistry from normal adult liver (AL) with negative hepatocytes and positive cholangiocytes (F), BA with strong immunoreactivity in hepatocyte nuclei (G), and BA-post-PE with less immunoreactivity in hepatocytes compared with BA (H). I: Western blotting of GATA6 protein in AL, BA, BA-post-PE, and disease control (DC) samples. NBI, normalized band intensity. J: paired sample analysis of GATA6 protein expression from patients before BA and after PE (BA-post-PE). Green (in situ hybridization) and brown (immunohistochemistry) indicate positive staining. Scale bars = 150 µm (B and C), 20 µm (C and E), and 200 µm (F and H).

Article Snippet: Primary antibodies used were as follows: rabbit antihuman GATA6 IgG (sc-9055; Santa Cruz Biotechnologies, Santa Cruz, CA) at dilution 1:1,200 for human samples, goat anti-human GATA6 IgG (AF1700; R&D Systems, Minneapolis, MN) at dilution 1:1,200 for mouse samples.

Techniques: Expressing, In Situ Hybridization, Immunohistochemistry, Western Blot, Staining

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Transcription factor GATA6: a novel marker and putative inducer of ductal metaplasia in biliary atresia

doi: 10.1152/ajpgi.00362.2017

Figure Lengend Snippet: P values from comparisons of mRNA levels of different liver sample types (in Figs. 2A and 7A, C, and E) using Wilcoxon method

Article Snippet: Primary antibodies used were as follows: rabbit antihuman GATA6 IgG (sc-9055; Santa Cruz Biotechnologies, Santa Cruz, CA) at dilution 1:1,200 for human samples, goat anti-human GATA6 IgG (AF1700; R&D Systems, Minneapolis, MN) at dilution 1:1,200 for mouse samples.

Techniques:

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Transcription factor GATA6: a novel marker and putative inducer of ductal metaplasia in biliary atresia

doi: 10.1152/ajpgi.00362.2017

Figure Lengend Snippet: GATA6 immunoreactivity in liver sample groups

Article Snippet: Primary antibodies used were as follows: rabbit antihuman GATA6 IgG (sc-9055; Santa Cruz Biotechnologies, Santa Cruz, CA) at dilution 1:1,200 for human samples, goat anti-human GATA6 IgG (AF1700; R&D Systems, Minneapolis, MN) at dilution 1:1,200 for mouse samples.

Techniques: Expressing

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Transcription factor GATA6: a novel marker and putative inducer of ductal metaplasia in biliary atresia

doi: 10.1152/ajpgi.00362.2017

Figure Lengend Snippet: Double in situ hybridization in biliary atresia (BA) liver. A and B: double in situ hybridization was performed on 2 different samples of BA liver. GATA6 expression is high in both ductular reaction (DR) area and liver parenchyma (hepatocytes); these 2 histological compartments are denoted in the figures by dashed lines. C and D: CK7 (C) is strongly expressed in DR area, whereas its expression is weaker in hepatocytes. CFTR (D) expression is limited to DR area. E and F: merged images of DAPI, GATA6, and CK7 (E) or DAPI, GATA6, and CFTR (F). Bile duct epithelium (arrow) is positive for both GATA6 and CK7/CFTR, but hepatocytes (arrowhead) express only GATA6. G and I: higher-magnification images from E. H and J: higher-magnification images from F. Green indicates positive signal for GATA6, and red indicates positive signal for CK7 or CFTR. Blue indicates DAPI staining in the nuclei of all cell types. Scale bars = 50 µm (A–F) and 10 µm (G–J).

Article Snippet: Primary antibodies used were as follows: rabbit antihuman GATA6 IgG (sc-9055; Santa Cruz Biotechnologies, Santa Cruz, CA) at dilution 1:1,200 for human samples, goat anti-human GATA6 IgG (AF1700; R&D Systems, Minneapolis, MN) at dilution 1:1,200 for mouse samples.

Techniques: In Situ Hybridization, Expressing, Staining

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Transcription factor GATA6: a novel marker and putative inducer of ductal metaplasia in biliary atresia

doi: 10.1152/ajpgi.00362.2017

Figure Lengend Snippet: GATA6 is overexpressed in hepatocytes in 2 mouse models of biliary obstruction. A: GATA6 protein is expressed in bile duct epithelium (arrow) but not in hepatocytes (arrowhead) of normal murine liver (4 wk after sham surgery). B: in mice subjected to bile duct ligation (BDL), hepatocytes (arrowhead) in the periportal area strongly express GATA6. C and D: at postnatal day 15 (P15), before the onset of ductular reaction (DR), there remains weak expression of GATA6 in normal hepatocytes (arrowhead), and there is no difference in GATA6 expression between control (Ctrl) and Alb-Cre;Rbpjflox/flox;Hnf6flox/flox double knockout (DKO) mice. E and F: at P30 Ctrl hepatocytes are GATA6 negative, whereas the Alb-Cre;Rbpjflox/flox;Hnf6flox/flox liver with severe DR shows strong GATA6 immunoreactivity in hepatocytes. G and H: at P120 in Alb-Cre;Rbpjflox/flox;Hnf6flox/flox liver, the DR has diminished, and liver histology, as well as GATA6 expression, is similar to Ctrl liver with immunoreactivity only in bile duct epithelial cells (arrow). n = 3 in each group. Scale bars = 50 µm.

Article Snippet: Primary antibodies used were as follows: rabbit antihuman GATA6 IgG (sc-9055; Santa Cruz Biotechnologies, Santa Cruz, CA) at dilution 1:1,200 for human samples, goat anti-human GATA6 IgG (AF1700; R&D Systems, Minneapolis, MN) at dilution 1:1,200 for mouse samples.

Techniques: Ligation, Expressing, Double Knockout

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Transcription factor GATA6: a novel marker and putative inducer of ductal metaplasia in biliary atresia

doi: 10.1152/ajpgi.00362.2017

Figure Lengend Snippet: In patients with biliary atresia (BA), GATA6 protein expression in hepatocytes correlates to bile duct expansion (BDE), age at portoenterostomy (PE), and the liver injury marker alanine aminotransferase. Hepatocyte GATA6 protein expression was divided into 2 groups (<70% positive nuclei = low/intermediate vs. >70% positive nuclei = high) and correlated to BDE rate (P = 0.0094) (A), age at PE (B), and plasma (P)-alanine aminotransferase (C). A: contingency tabling coupled with χ2 test was employed to test the statistical significance. B and C: dots represent individual samples, the box represents the interquartile range, and the whiskers represent the 1st and 4th quartile. The line inside the box is the median, and the dashed line represents the mean. *P < 0.05.

Article Snippet: Primary antibodies used were as follows: rabbit antihuman GATA6 IgG (sc-9055; Santa Cruz Biotechnologies, Santa Cruz, CA) at dilution 1:1,200 for human samples, goat anti-human GATA6 IgG (AF1700; R&D Systems, Minneapolis, MN) at dilution 1:1,200 for mouse samples.

Techniques: Expressing, Marker

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Transcription factor GATA6: a novel marker and putative inducer of ductal metaplasia in biliary atresia

doi: 10.1152/ajpgi.00362.2017

Figure Lengend Snippet: Enforced expression of GATA6 causes alterations in expression of genes regulating cholangiocyte and hepatocyte differentiation. The human hepatocellular cell line HepG2 and primary human hepatocytes were transiently transfected with pCDNA3-GATA6 or pCDNA3 plasmid alone. After 48 h, RNA was harvested and subjected to qRT-PCR analysis. A: in HepG2 cells, 3 genes related to cholangiocyte differentiation (HNF1β, HNF6, and JAG1) were significantly upregulated in cells overexpressing GATA6 compared with control (Ctrl) cells. B: in primary human hepatocytes, the relative expression of 4 genes related to cholangiocyte differentiation (HNF1β, HNF6, JAG1, and DKK1) and 1 gene related to hepatocyte differentiation (HNF4α) were upregulated in cells with GATA6 overexpression compared with control cells. For both cell types, 5 independent experiments were performed in triplicate. Each bar depicts the logarithm of the ratio of mRNA expression GATA6 overexpression vs. control cells. *P < 0.05, **P < 0.01.

Article Snippet: Primary antibodies used were as follows: rabbit antihuman GATA6 IgG (sc-9055; Santa Cruz Biotechnologies, Santa Cruz, CA) at dilution 1:1,200 for human samples, goat anti-human GATA6 IgG (AF1700; R&D Systems, Minneapolis, MN) at dilution 1:1,200 for mouse samples.

Techniques: Expressing, Transfection, Plasmid Preparation, Quantitative RT-PCR, Over Expression

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Transcription factor GATA6: a novel marker and putative inducer of ductal metaplasia in biliary atresia

doi: 10.1152/ajpgi.00362.2017

Figure Lengend Snippet: Expression of HNF1β, HNF6, and JAG1 is elevated in biliary atresia (BA) liver, decreases after successful portoenterostomy (PE), and correlates to GATA6. A, C, and E: boxplots showing relative mRNA expression of HNF1β (A), HNF6 (C), and JAG1 (E), as measured by qPCR, in BA vs. other liver specimens. Dots represent individual samples, the box represents the interquartile range, and the whiskers represent the 1st and 4th quartile. The line inside the box is the median, and the dashed line represents the mean. *P < 0.05, **P < 0.01. The P values of other paired comparisons are indicated in Table 2. B, D, and F: linear regression analyses of HNF1β (B), HNF6 (D), and JAG1 (F) mRNA expression vs. GATA6 mRNA expression in BA-post-PE samples. FL, fetal liver; AL, adult liver; DC, disease control.

Article Snippet: Primary antibodies used were as follows: rabbit antihuman GATA6 IgG (sc-9055; Santa Cruz Biotechnologies, Santa Cruz, CA) at dilution 1:1,200 for human samples, goat anti-human GATA6 IgG (AF1700; R&D Systems, Minneapolis, MN) at dilution 1:1,200 for mouse samples.

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Development of Human Adrenocortical Adenoma (HAA1) Cell Line from Zona Reticularis

doi: 10.3390/ijms24010584

Figure Lengend Snippet: ( A ) HAA1 cells in culture. ( B ) Both master-regulator NR5A1/SF1 (green) and the key adrenal transcription GATA4 and GATA6 proteins (red) are expressed at a lower level in the HAA1 cell line (bottom panels) than in the control NCI-H295R (top panels) human adrenal cortical carcinoma cell line. Scale bar, 50 µ. ( C ) A representative Western blot analysis of protein expression; NR5A1/SF1 (top panel) and ACTIN1 (bottom panel); L1. SW13 adrenal carcinoma cells (these cells produce no steroids and serve as a negative control) L2. NCI-H295.L3. HAA1 ( D ) Quantitative analysis of GATA4 and GATA6 protein expression for the experiment shown in ( B ); *, p < 05; ***, p < 0.001. ( E ) Quantitative analysis of NR5A1/SF1 expression for the experiment shown in ( C ). All differences are significant, p < 0.05.

Article Snippet: The following antibody combinations were used: goat anti-GATA4 antibody (R&D Systems, Minneapolis, MN, USA), followed by donkey anti-goat Alexa Fluor 555-conjugated antibodies (Alexa); rabbit anti-GATA6 antibody (Cell Signaling Technology, Danvers, MA, USA) and rabbit anti-GLI1 antibody (Santa Cruz Biotechnology, Dallas, TX, USA) followed by goat anti-rabbit Alexa Fluor 488-conjugated antibodies (Invitrogen); and goat anti-SHH antibody, goat anti-HSD3B2, and goat anti-CYP11A1 (all Santa Cruz Biotechnology) antibodies followed by donkey anti-goat Alexa Fluor 555-conjugated antibodies (Invitrogen).

Techniques: Control, Western Blot, Expressing, Negative Control