Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Inflammation Induces Lipid Deposition in Kidneys by Downregulating Renal PCSK9 in Mice with Adriamycin-Induced Nephropathy

doi: 10.12659/MSM.917312

Figure Lengend Snippet: ( A–F ) Renal expression of TGF-β1, IL-1β, HNF1α, PCSK9, LDLR, and SREBP2 proteins (immunohistochemical analysis software was used to analyze the optical density values in the images, and semi-quantitative values of positive expression in kidney tissues were obtained) and mRNAs (from RT-qPCR, relative to GAPDH) in the 4 groups at 4, 8, and 12 weeks. Values represent means ± standard errors of mean for groups of 8 mice each. For comparisons at the same time point: * P <0.05 versus CTL, # P <0.05 versus AWI. TGF-β1 – transforming growth factor beta 1; IL-1β – interleukin 1β; HNF1α – hepatocyte nuclear factor 1α; PCSK9 – pro-protein convertase subtilisin kexin type 9; LDLR – low-density lipoprotein receptor; SREBP2 – sterol regulatory element binding protein 2; RT-q-PCR – real-time quantitative polymerase chain reaction; GAPDH – glyceraldehyde 3-phosphate dehydrogenase; CLT – control group; AWI group – Adriamycin-induced nephrosis with inflammation group.

Article Snippet: Immunohistochemistry (IHC) was used to examine the expression of interleukin (IL)-1β, transforming growth factor beta 1 (TGF-β1), HNF1α, PCSK9, LDLR, and SREBP2 using the following antibodies: rabbit anti-rat IL-1β antibody (working dilution 1: 100; sc-7884, Santa Cruz Biotechnology, USA), rabbit anti-rat TGF-β1 (working dilution 1: 100; sc-146, Santa Cruz Biotechnology), mouse anti-human SREBP2 antibody (working dilution 1: 100; sc-271615, Santa Cruz Biotechnology), mouse anti-human HNF1 α antibody (working dilution 1: 100; sc-393668, Santa Cruz Biotechnology,), rabbit anti-rat LDLR antibody (working dilution 1: 100; ab30532, Abcam, UK), rabbit anti-rat PCSK9 antibody (working dilution 1: 100; ab31762, Abcam).

Techniques: Expressing, Immunohistochemical staining, Software, Quantitative RT-PCR, Binding Assay, Real-time Polymerase Chain Reaction, Control

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Inflammation Induces Lipid Deposition in Kidneys by Downregulating Renal PCSK9 in Mice with Adriamycin-Induced Nephropathy

doi: 10.12659/MSM.917312

Figure Lengend Snippet: Representative immunostaining results of renal tissues for HNF1α, PCSK9, LDLR, and SREBP2 in each group at 4, 8, and 12 weeks. Scale bars indicate 100 μm. HNF1α – hepatocyte nuclear factor 1α; PCSK9 – pro-protein convertase subtilisin kexin type 9; LDLR – low-density lipoprotein receptor; SREBP2 – sterol regulatory element binding protein 2.

Article Snippet: Immunohistochemistry (IHC) was used to examine the expression of interleukin (IL)-1β, transforming growth factor beta 1 (TGF-β1), HNF1α, PCSK9, LDLR, and SREBP2 using the following antibodies: rabbit anti-rat IL-1β antibody (working dilution 1: 100; sc-7884, Santa Cruz Biotechnology, USA), rabbit anti-rat TGF-β1 (working dilution 1: 100; sc-146, Santa Cruz Biotechnology), mouse anti-human SREBP2 antibody (working dilution 1: 100; sc-271615, Santa Cruz Biotechnology), mouse anti-human HNF1 α antibody (working dilution 1: 100; sc-393668, Santa Cruz Biotechnology,), rabbit anti-rat LDLR antibody (working dilution 1: 100; ab30532, Abcam, UK), rabbit anti-rat PCSK9 antibody (working dilution 1: 100; ab31762, Abcam).

Techniques: Immunostaining, Binding Assay

Journal: Cell Research

Article Title: Tissue-specific transcription reprogramming promotes liver metastasis of colorectal cancer

doi: 10.1038/s41422-019-0259-z

Figure Lengend Snippet: HNF1A is required and sufficient for the expression of liver specific genes in CRC cells and liver colonization and metastasis. a The blue and red bars in the chart show HNF1A gene expression in normal colon and liver tissues, respectively. The Y axis shows the mean RPKM value. The RNA-seq data were obtained from the GTEx project database. b UCSC genome browser images of an enhancer around the liver-specific gene HNF1A in SW620 and SW480 covered genomic regions marked by H3K27ac and H3K4me2. c Immunoblot analysis was used to assess efficiency of HNF1A knockdown in SW620 cells. GAPDH served as a loading control (Top panel). GSEA of liver-specific signatures as defined from the GTEx project database is shown. Genes are ranked by the log2 fold change of the FPKM values in HNF1A-knockdown versus shNC cells. The NES and FDR are shown (Bottom two panels). d Gene ontology analysis of down-regulated genes by HNF1A knockdown. e Immunoblot analysis was used to assess efficiency of HNF1A overexpression in HCT116 cells. GAPDH served as a loading control (Top panel). GSEA of liver-specific signatures as defined from the GTEx project database is shown. Genes are ranked by the log2 fold change of the FPKM values in HCT116 HNF1A-overexpression versus Control cells. The NES and FDR are shown (Bottom panel). f HNF1A is highly expressed in human liver metastatic CRC tumors as compared with primary tumors. The expression data are from the GSE41258 dataset. Each dot represents one primary or liver metastatic CRC tumor, and the red bar represents the mean value. Statistically-significant P values are indicated with asterisks (***P < 0.001, by t-test). g Luciferase measurements from different time points upon intracecal injection of HCC022 shNC and shHNF1A cells. An shHNF1A virus pool (3 HNF1A shRNA sequences) was expressed in HCC022 cells and shNC and shHNF1A cells were injected into mice using intracecal injection (n = 4 shNC mice and n = 4 shHNF1A mice). Then a time-course analysis of luciferase measurements in mice was performed; Black lines represent shNC control group and red lines represent shHNF1A mice group. Error bars represent standard deviation of the mean. Student’s t test, P < 0.01, n.s., not significant. h The stacked bars indicate the percentage of mice with liver metastases or with no liver metastases relative to the total number of mice in each section. The hepatic metastasis rate of HCC022 cells is indicated at the bottom. Data were analyzed using Pearson’s Chi-square test. **P < 0.01. i Representative images of the metastases upon intracecal injection of HCT116 control and HNF1A-overexpressing cells. j The stacked bars indicate the percentage of mice with liver metastases or with no liver metastases relative to the total number of mice in each section. The hepatic metastasis rate is indicated at the bottom. Data were analyzed using Pearson’s Chi-square test. *P < 0.05. k Representative images of the metastases upon intrahepatic injection of control and HNF1A-overexpressing HCT116 cells. l The stacked bars indicate the percentage of mice with liver colonies and without liver colonies relative to the total number of mice in each section. The hepatic colonization rate is indicated at the bottom. Data were analyzed using Pearson’s Chi-square test. **P < 0.01

Article Snippet: The following primary antibodies were used: anti-FOXA2 (Abcam Cat. No. ab108422), anti-HNF1A (proteintech Cat. No. 22426–1-AP), and anti-GAPDH (Cell Signaling Technology Cat. No. 2118) antibodies.

Techniques: Expressing, RNA Sequencing Assay, Western Blot, Over Expression, Luciferase, Injection, shRNA, Standard Deviation

Journal: The Journal of clinical endocrinology and metabolism

Article Title: Early development of hyperparathyroidism due to loss of PTH transcriptional repression in patients with HNF1β mutations?

doi: 10.1210/jc.2012-3453

Figure Lengend Snippet: Figure 1. HNF1 expression in human parathyroid glands. A, Endogenous expression of HNF1 in human parathyroid tissue was investigated using RT-PCR. PTH was included as positive control for tissue specificity and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as housekeeping gene. RT indicates RT sample; RT indicates no RT control; cDNA indicates no cDNA control. B, Representative images of immunohistochemical analysis of HNF1 and PTH colocalization in human parathyroid tissue. DAPI is a nuclear marker. Scale bars, 20 m.

Article Snippet: Samples were incubated with 5.0 g rabbit polyclonal antihuman HNF1 antibody (Santa Cruz; sc-22840), rabbit IgG isotype antibodies as negative control, or anti-trimethylhistone H3 antibodies as positive control, which were bound to protein A magnetic beads.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Positive Control, Control, Immunohistochemical staining, Marker

Journal: The Journal of clinical endocrinology and metabolism

Article Title: Early development of hyperparathyroidism due to loss of PTH transcriptional repression in patients with HNF1β mutations?

doi: 10.1210/jc.2012-3453

Figure Lengend Snippet: Figure 2. The human PTH gene as target of the HNF1 transcription factor. A, Sequence logo of the HNF1 motif from the Jaspar database (ID MA0153.1). In the human PTH promoter region (1476 bp from the transcription initiation site), 3 putative HNF1 binding sites map at 1238, 690, and 101 of the forward DNA strand (). B, Sequence alignment of the 3 putative HNF1 recognition sites in the PTH promoter region of different mammalian species. C, Binding of the PTH promoter by HNF1 was verified by a ChIP assay in HEK293 cells cotransfected with the human PTH promoter luciferase constructs, 1476/25 or 70/25, and wild-type HNF1. For immunoprecipitation, an anti-HNF1 antibody or a rabbit IgG isotype control antiserum was used. Immunoprecipitation with an anti-trimethyl-histone H3 antibody was included as positive control. Two percent of the chromatin used for immunoprecipitation (input) was also included as a control. D, Dose-response curve of increasing amounts of wild-type HNF1 cotransfected in HEK293 cells with a luciferase construct carrying the human PTH gene promoter region 1476/ 25 (n 9). **, P .001, compared with mock.

Article Snippet: Samples were incubated with 5.0 g rabbit polyclonal antihuman HNF1 antibody (Santa Cruz; sc-22840), rabbit IgG isotype antibodies as negative control, or anti-trimethylhistone H3 antibodies as positive control, which were bound to protein A magnetic beads.

Techniques: Sequencing, Binding Assay, Luciferase, Construct, Immunoprecipitation, Control, Positive Control

Journal: The Journal of clinical endocrinology and metabolism

Article Title: Early development of hyperparathyroidism due to loss of PTH transcriptional repression in patients with HNF1β mutations?

doi: 10.1210/jc.2012-3453

Figure Lengend Snippet: Figure 3. HNF1 mutants lack the ability to inhibit PTH promoter activity. A, Study of the human PTH promoter activity when cotransfected with wild-type HNF1 or HNF1 p.His69fsdelAC, p.Lys156Glu, p.His324Ser325fsdelCA, p.Tyr352fsinsA. mutants or the mock plasmid in HEK293 cells (n 3). **, P .001, compared with wild-type HNF1. B, Serial deletion analysis in which the PTH promoter activity was tested in HEK293 cells in the presence of wild-type HNF1 (empty bars) or HNF1 p.His324Ser325fsdelCA mutant (black bars) (n 9). **, P .001, compared with HNF1 p.His324Ser325fsdelCA; *, P .006, compared with HNF1 p.His324Ser325fsdelCA; ns, nonsignificant compared with HNF1 p.His324Ser325fsdelCA, P .9.

Article Snippet: Samples were incubated with 5.0 g rabbit polyclonal antihuman HNF1 antibody (Santa Cruz; sc-22840), rabbit IgG isotype antibodies as negative control, or anti-trimethylhistone H3 antibodies as positive control, which were bound to protein A magnetic beads.

Techniques: Activity Assay, Plasmid Preparation, Mutagenesis

Journal: iScience

Article Title: Suppression of preadipocyte determination by SOX4 limits white adipocyte hyperplasia in obesity

doi: 10.1016/j.isci.2023.106289

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti--TCF1 , Proteintech , Cat#14464-1-AP; RRID: AB_2878061.

Techniques: Virus, Recombinant, Lysis, BIA-KA, shRNA, Luciferase, Plasmid Preparation, Software, Magnetic Beads

Journal: Hormones (Athens, Greece)

Article Title: HNF1A gene mutations and premature ovarian failure (POF): evidence from a clinical paradigm combining MODY 3 and POF

doi: 10.1007/s42000-024-00529-y

Figure Lengend Snippet: Immunohistochemical staining indicating HNF1A expression in fetal ovaries (HNF1A X 400). A No expression at 23 weeks of gestation. B , C Positive expression in the primordial follicles at 32 and 40 weeks of gestation

Article Snippet: Expression of the HNF1A protein was examined by immunohistochemistry using a rabbit polyclonal antibody that binds to human HNF1A (HNF1 NBP1-33,596 Novus Biologicals, diluted 1:200, pretreated with citrate buffer pH = 6, prior to primary antibody addition).

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Hormones (Athens, Greece)

Article Title: HNF1A gene mutations and premature ovarian failure (POF): evidence from a clinical paradigm combining MODY 3 and POF

doi: 10.1007/s42000-024-00529-y

Figure Lengend Snippet: Immunohistochemical staining indicating HNF1A protein expression in the ovaries of adult premenopausal women. A Primary ovarian follicles (× 200). B Corpus luteum of menstruation: faint HNF1A expression in the thick inner layer of large granulosa-lutein cells and more intense expression in the outer thinner layer of smaller theca-lutein cells (× 100)—between arrow heads. C Luteinized stromal cells (× 200). D Epithelial inclusion glands of the ovarian cortex (× 200)

Article Snippet: Expression of the HNF1A protein was examined by immunohistochemistry using a rabbit polyclonal antibody that binds to human HNF1A (HNF1 NBP1-33,596 Novus Biologicals, diluted 1:200, pretreated with citrate buffer pH = 6, prior to primary antibody addition).

Techniques: Immunohistochemical staining, Staining, Expressing