Journal: Scientific Reports

Article Title: A common genetic variation of melanoma inhibitory activity-2 labels a subtype of pancreatic adenocarcinoma with high endoplasmic reticulum stress levels

doi: 10.1038/srep08109

Figure Lengend Snippet: (a), Pancreatic islet cells are immunopositive for HNF1A and MIA2 (scale bar: 100 μm). (b), 73% and 52% of the samples are positive for HNF1A and MIA2, respectively (scale bar: 100 μm). (c), Protein expression of HNF1A and MIA2 in pancreatic cancer cell lines. Out of 7 tested cell lines (Aspc-1 (As), Bxpc-3 (Bx), Colo357 (Co), Mia-PaCa2 (Mi), Panc-1 (Pa), T3M4 (T3), Su86.86 (Su)), Aspc-1 and Colo-357 express HNF1A and MIA2; endogenous MIA2 is mainly detected at a size of 70 kDa (loading control: β-actin). (d), MRNA expression of MIA2 in 7 pancreatic cancer cell lines. (e), MIA2 silencing with two sets of specific siRNAs (#1 and #2) or negative control siRNA (control) at 72 h in Aspc-1 cells and Colo-357 cells; results of densitometry are shown on top of the blot. One representative blot out of three independent experiments is shown. (f), Transient expression of HNF1A in Su86.86 and Panc-1 cells by transfection of an HNF1A expression vector (upper panel), loading control: GAPDH; MIA2 mRNA expression is induced by transient expression of HNF1A in Su86.86 and Panc-1 cells. HPRT1: housekeeping gene. One of three independent experiments is shown. The gels were run under the same experimental conditions and cropped blots/gels are presented (full-length blots/gels are provided in with indicated cropping lines).

Article Snippet: For transient expression of HNF1A, the expression vector from Origene was used (RC211201, Origene, Suite 200 Rockville, MD).

Techniques: Expressing, Control, Negative Control, Transfection, Plasmid Preparation

Journal: Cells

Article Title: Dual Role of HNF4α in Colorectal Adenocarcinoma During Carcinogenesis and Metastasis.

doi: 10.3390/cells14080599

Figure Lengend Snippet: Figure 1. (a) Differential expression of HNF4α from RNA sequencing data of TCGA samples. Higher HNF4α expression was detected in colon and rectal AC (adenocarcinoma) compared to HCC. (b) HNF4α expression in CRAC tissues and their matched normal counterparts (* p < 0.01). (c) Protein expression of HNF4α among different cancer types from Human Protein Atlas data samples. Protein expression of HNF4α was the highest in CRAC compared to STAD and PAAD. (d) Immunoblot analyses of HNF4α (P1/P2) in colon cancer cell lines. Strong HNF4α expression in the LoVo, HCT116, and Caco2 cell lines, while weak or minimal HNF4α expression in the SW480, COLO205, and HT29 cell lines observed in the Western blot.

Article Snippet: The pCMV6-Entry vector (PS100001) and human HNF4α cDNA clone (RC217863) were purchased from Origene Technologies, Inc. (Rockville, MD, USA).

Techniques: Quantitative Proteomics, RNA Sequencing, Expressing, Western Blot

Journal: Cells

Article Title: Dual Role of HNF4α in Colorectal Adenocarcinoma During Carcinogenesis and Metastasis.

doi: 10.3390/cells14080599

Figure Lengend Snippet: Figure 2. (a) SW480 cells expressing HNF4α displayed increased migration compared to their control counterparts. (b) Assessment of the migration and invasion abilities of LoVo and Caco2 cells with HNF4α inhibition using the Transwell assay. (c) Wound-healing assay of LoVo and COLO205 cells, indicating a decrease in the percentage of wound closure and migration rate in HFN4α-inhibited LoVo cells.

Article Snippet: The pCMV6-Entry vector (PS100001) and human HNF4α cDNA clone (RC217863) were purchased from Origene Technologies, Inc. (Rockville, MD, USA).

Techniques: Expressing, Migration, Control, Inhibition, Transwell Assay, Wound Healing Assay

Journal: Cells

Article Title: Dual Role of HNF4α in Colorectal Adenocarcinoma During Carcinogenesis and Metastasis.

doi: 10.3390/cells14080599

Figure Lengend Snippet: Figure 3. Immunohistochemical localization of HNF4α in human tissue samples of (a) normal colonic epithelia, (b) tubular adenoma (left) and primary CRAC (right), (c) metastatic CRAC in the liver, (d) moderately differentiated CRAC, (e) poorly differentiated CRAC, and (f) poorly cohesive CRAC. (g) Primary CRAC samples displayed higher HNF4α expression than normal epithelia and metastatic CRAC (p = 0.002 and p = 0.012). (h) Well- and moderately differentiated CRAC samples displayed elevated expression of HNF4α. (i) Immunoblot assay and immunohistochemical analyses of primary and metastatic (synchronous and metachronous) CRAC (i–m). Synchronous metastatic (Mets) CRAC displayed lower expression and metachronous Mets displayed higher levels of HNF4α compared to primary CRAC. Scale bar = 100 µm.

Article Snippet: The pCMV6-Entry vector (PS100001) and human HNF4α cDNA clone (RC217863) were purchased from Origene Technologies, Inc. (Rockville, MD, USA).

Techniques: Immunohistochemical staining, Expressing, Western Blot

Journal: Cells

Article Title: Dual Role of HNF4α in Colorectal Adenocarcinoma During Carcinogenesis and Metastasis.

doi: 10.3390/cells14080599

Figure Lengend Snippet: Figure 4. Kaplan–Meier survival curves of HNF4α expression in CRAC. (a) Determination of cutoff values (red lines) using the cutoff finder. (b) Overall and progression-free survival curves obtained with data from HNF4α mRNA level from the TCGA (p = 0.022 and p = 0.033). (c) Overall and disease-free survival curves obtained using HNF4α protein level data from CNUH (p = 0.017 and p = 0.021).

Article Snippet: The pCMV6-Entry vector (PS100001) and human HNF4α cDNA clone (RC217863) were purchased from Origene Technologies, Inc. (Rockville, MD, USA).

Techniques: Expressing

Journal: Cells

Article Title: Dual Role of HNF4α in Colorectal Adenocarcinoma During Carcinogenesis and Metastasis.

doi: 10.3390/cells14080599

Figure Lengend Snippet: Figure 5. Immunoblot assay of the NF-κB, YAP, and EMT markers in CRAC cell lines (a) Decrease in NF-κB markers under conditions of HNF4α inhibition in LoVo and Caco2 cells. (b) Increase in YAP, E-cadherin, and N-cadherin under conditions of HNF4α inhibition in LoVo and Caco2 cells. Decrease in EpCAM and Vimentin and, conversely, increase in EpCAM and Vimentin in LoVo cells depleted of HNF4α. (c) Immunohistochemical changes in pNF-κb and YAP in tubular adenoma, primary CRAC, and metastatic CRAC.

Article Snippet: The pCMV6-Entry vector (PS100001) and human HNF4α cDNA clone (RC217863) were purchased from Origene Technologies, Inc. (Rockville, MD, USA).

Techniques: Western Blot, Inhibition, Immunohistochemical staining

Journal: Cells

Article Title: Dual Role of HNF4α in Colorectal Adenocarcinoma During Carcinogenesis and Metastasis.

doi: 10.3390/cells14080599

Figure Lengend Snippet: Figure 6. Hypothesis of HNF4α in relation to signaling pathways: low HNF4α state related to the suppression of NF-κb pathways, an increase in epithelial markers, and an increase in YAP levels. Changes in HNF4α expression in CRAC patients: (i) increased HNF4α expression during early carcinogenesis, (ii) lower expression during progression and dedifferentiation, and (iii) higher expression in metachronous metastasis.

Article Snippet: The pCMV6-Entry vector (PS100001) and human HNF4α cDNA clone (RC217863) were purchased from Origene Technologies, Inc. (Rockville, MD, USA).

Techniques: Protein-Protein interactions, Expressing