Journal: OncoTargets and therapy

Article Title: Carvacrol suppresses proliferation and invasion in human oral squamous cell carcinoma

doi: 10.2147/OTT.S98875

Figure Lengend Snippet: Carvacrol inhibits Tca-8113 cell migration and invasion. Notes: ( A ) The expression of p-FAK and FAK proteins was analyzed by Western blotting in carvacrol-treated Tca-8113 cells. ( B , C ) Expression of β-catenin and ZEB1 in carvacrol-treated Tca-8113 cells. Then, Tca-8113 cells were treated by control or 40 μM of carvacrol for 24 hours, respectively. ( D ) Transwell assay analyses of the migration or invasion potential of Tca-8113 cells. Magnification ×200. ( E ) Numbers of migratory or invasive cells (mean ± standard deviation, n=5, “n” means five randomly chosen fields). ** P <0.01. ( F , G ) Tca-8113 cells were treated with 0 to 80 μM of carvacrol for 24 hours. MMP-2 and MMP-9 levels were determined by Western blotting.

Article Snippet: P21, CDK2, CDK4, CDK6, CCND1, Cox2, Bcl-2, Bax, FAK, p-FAK, ZEB1, and β-catenin were purchased from Cell Signaling Technology (Danvers, MA, USA). β-actin was purchased from Beijing Zhongshan Goldenbridge Biotechnology (Beijing, People’s Republic of China) and GAPDH was purchased from ShangHai Kangchen (Shanghai, People’s Republic of China).

Techniques: Migration, Expressing, Western Blot, Transwell Assay, Standard Deviation

Journal: International Journal of Biological Sciences

Article Title: Reactive Oxygen Species Are Involved in Regulating Hypocontractility of Mesenteric Artery to Norepinephrine in Cirrhotic Rats with Portal Hypertension

doi: 10.7150/ijbs.8081

Figure Lengend Snippet: Expression and interaction of α1 adrenergic receptor and β-arrestin-2. Western blot was used to determine the expression of α1 adrenergic receptor and β-arrestin-2. Co-immunoprecipitation was performed to investigate the interaction between α1 adrenergic receptor and β-arrestin-2. ( A ) α1 adrenergic receptor and β-arrestin-2 expression in all the groups. ( B ) Quantitative analysis of β-arrestin-2 expression. The bar graphs show the relative expression levels of indicated proteins after normalization to β-actin. Values are mean ± SEM; n=7 for each group. ( C ) Interaction between the α1 adrenergic receptor and β-arrestin-2 in mesenteric artery of the rats. n = 7 for each group. **: compared with NC, P <0.01; #: compared with PHTC, P <0.05; ##: compared with PHTC, P <0.01.

Article Snippet: The following antibodies were used in the study: Rabbit anti-ROCK-1 antibody (CST, American), normal rabbit IgG (sc-2027), rabbit anti-mosein antibody (sc-6410), rabbit anti-phospho-moesin threonine 555 (sc-12895), mouse anti-β-arrestin-2 antibody (sc-6387), rabbit anti-α1 adrenergic receptor antibody.

Techniques: Expressing, Western Blot, Immunoprecipitation

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Deficiency of Stomach-Type Claudin-18 in Mice Induces Gastric Tumor Formation Independent of H pylori Infection

doi: 10.1016/j.jcmgh.2019.03.003

Figure Lengend Snippet: Activation of inflammatory signals including CCL28 and CXCL5 in aged stCldn18–/– mice. ( A ) Expression levels of IL-1β, TNF-α, and CXCL1 in the stomach from stCldn18+/+ (n = 8, 4, 11, 10) and stCldn18–/– mice (n = 14, 4, 13, 16) (8, 40, 60, and 100 w.o.) quantified by qRT-PCR. Gene expressions were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Results are expressed as mean ± SD. Comparisons between 2 groups were performed using Student’s t test, and differences with P < .05 were considered statistically significant. * P < .05; ** P < .01. ( B ) Immunofluorescence micrographs for CXCL5 ( green ) co-stained with α-catenin ( red ) and DAPI ( blue ) in the stomach from stCldn18+/+ mice and stCldn18–/– mice (8, 40, 60, and 100 w.o.). CXCL5 signals were clearly observed in the stomach of stCldn18–/– mice after middle-age (40, 60, and 100 w.o.) ( arrows ). High-magnification images are shown in insets. Representative images from at least 3 independent experiments are shown. Scale bars = low magnification: 50 μm, high magnification: 10 μm. ( Right ) Expression level of CXCL5 in the stomach from stCldn18+/+ (n = 8, 4, 12, 10) and stCldn18–/– mice (n = 6, 4, 13 16) (8, 40, 60, and 100 w.o.) quantified by qRT-PCR. Gene expressions were normalized to GAPDH. Results are expressed as mean ± SD. Comparisons between 2 groups were performed using Student’s t test, and differences with P < .05 were considered statistically significant. ** P < .01. ( C ) Expression levels of STAT3, IKKβ, and NF-κB in the stomach from stCldn18+/+ (n = 4, 4, 10, 10) and stCldn18–/– mice (n = 6, 4, 12, 16) (8, 40, 60, and 100 w.o.) quantified by qRT-PCR. Gene expressions were normalized to GAPDH. Results are expressed as mean ± SD. Comparisons between 2 groups were performed using Student’s t test, and differences with P < .05 were considered statistically significant. * P < .05; ** P < .01. ( D ) ( Left ) Immunofluorescence micrographs for CCL28 ( green ), a lymphocyte chemoattractant and maintainer of chronic inflammation, co-stained with α-catenin ( red ) and DAPI ( blue ) in the stomach from stCldn18+/+ and stCldn18–/– mice (10, 40, 60, and 100 w.o.). CCL28 signals were clearly observed in the stomach of the stCldn18–/– mice after middle-age (40, 60, and 100 w.o.) ( arrows ). High-magnification images are shown in insets. Representative images from at least 3 independent experiments are shown. Scale bars = low magnification: 50 μm, high magnification: 10 μm. ( Right ) Expression level of CCL28 in the stomach from stCldn18+/+ (n = 8, 4, 12, 10) and stCldn18–/– (n = 10, 4, 13 16) mice (8, 40, 60, and 100 w.o.) quantified by qRT-PCR. Gene expressions were normalized to GAPDH. Results are expressed as mean ± SD. Comparisons between 2 groups were performed using Student’s t test, and differences with P < .05 were considered statistically significant. ** P < .01. ( E ) Expression levels of CXCL5, IKKβ, NFκB, and ASCL2 in the stomach from stCldn18+/+ and stCldn18–/– mice (around 50 w.o.) after POL7085 or cimetidine treatment, quantified by qRT-PCR (n: stCldn18–/– = 7, 5, 6, given saline, POL7085, and cimetidine, respectively). Gene expressions were normalized to GAPDH. Results are expressed as mean ± SD. Comparisons between 2 groups were performed using Student’s t test, and differences with P < .05 were considered statistically significant. * P < .05; ** P < .01.

Article Snippet: Active β-catenin , Cell Signaling Technology , 88145 , Unconjugate.

Techniques: Activation Assay, Expressing, Quantitative RT-PCR, Immunofluorescence, Staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Deficiency of Stomach-Type Claudin-18 in Mice Induces Gastric Tumor Formation Independent of H pylori Infection

doi: 10.1016/j.jcmgh.2019.03.003

Figure Lengend Snippet: Upregulation of TLR2 in aged stCldn18–/– mice. (A) ( Left ) Immunofluorescence micrographs for TLR2 ( green ) co-stained with α-catenin ( red ) and DAPI ( blue ) in the stomach from stCldn18+/+ and stCldn18–/– mice (8, 40, 60, and 100 w.o.). High-magnification images are shown in insets. TLR2-positive cells were clearly observed in the stomach of stCldn18–/– mice after middle age (40, 60, and 100 w.o.). At middle age (40 w.o.), the TLR2-positive cells did not show the characteristics of gastric epithelium ( arrowheads ), but at old age (60, and 100 w.o.), the TLR2-positive cells might have been gastric epithelial cells ( arrows ). Representative images from at least 2 independent experiments are shown. Scale bars = low magnification: 50 μm, high magnification: 50 μm. ( Right ) Expression level of TLR2 in the stomach from stCldn18+/+ (n = 4, 4, 10, 10) and stCldn18–/– mice (n = 8, 4, 12, 16) (8, 40, 60, and 100 w.o.) quantified by qRT-PCR. Results are expressed as mean ± SD. Comparisons between 2 groups were performed using Student’s t test, and differences with P < .05 were considered statistically significant. ** P < .01. ( B ) Immunofluorescence micrographs for CD44 ( green ), co-stained with TLR2 ( red ) in the stomach from stCldn18+/+ and stCldn18–/– mice (100 w.o.). High-magnification images are shown in insets. CD44-and TLR2-double-positive cells were clearly observed in the stomach of the stCldn18–/– mice ( arrows ). Representative images from at least 3 independent experiments are shown. Scale bars = low magnification: 50 μm, high magnification: 50 μm. (C) Expression levels of c-Myc, hypoxia-inducible factor HIF2a, MMP7, SOX2, and ASCL2 in the stomach from stCldn18+/+ (n = 5, 4, 12, 9) and stCldn18–/– (n = 9, 4, 13, 16) mice (8, 40, 60, and 100 w.o.) quantified by qRT-PCR. The expression level of MMP7 was especially increased in the stomach of the 100 w.o. stCldn18–/– mice. Gene expressions were normalized to GAPDH. Results are expressed as mean ± SD. Comparisons between 2 groups were performed using Student’s t test, and differences with P < .05 were considered statistically significant. * P < .05; *** P < .001.

Article Snippet: Active β-catenin , Cell Signaling Technology , 88145 , Unconjugate.

Techniques: Immunofluorescence, Staining, Expressing, Quantitative RT-PCR

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Deficiency of Stomach-Type Claudin-18 in Mice Induces Gastric Tumor Formation Independent of H pylori Infection

doi: 10.1016/j.jcmgh.2019.03.003

Figure Lengend Snippet: Activation of Wnt signaling in aged stCldn18–/– mice. ( A ) Immunohistological micrographs for β-catenin as an EMT marker in the stomach from stCldn18+/+ and stCldn18–/– mice (8, 40, 60, and 100 w.o.). A nuclear localization of β-catenin, indicative of Wnt signaling activation, was observed in the stomach of stCldn18 –/– mice after middle age (40, 60, and 100 w.o.) ( arrows ). Representative images from at least 3 independent experiments are shown. Scale bars = 50 μm. ( B ) Expression level of Wnt1a in the stomach from stCldn18+/+ mice (n = 11, 10), and stCldn18–/– mice with hyperplasia (n = 8, 13), with gastric tumor (n = 5, 3) (60, and 100 w.o.) quantified by qRT-PCR. Wnt1a was especially increased in the gastric tumors of the 100-w.o. stCldn18–/– mice. Gene expressions were normalized to GAPDH. ( C ) Expression level of Wnt2 , Wnt3 , Wnt4 , and Wnt5 in the stomach from stCldn18+/+ (n = 5, 4, 12, 9) and stCldn18–/– (n = 9, 4, 13, 16) mice (10, 40, 60, and 100 w.o.), quantified by qRT-PCR. Gene expressions were normalized to GAPDH. Results are expressed as mean ± SD. Comparisons between 2 groups were performed using Student’s t test, and differences with P < .05 were considered statistically significant. * P < .05; *** P < .001.

Article Snippet: Active β-catenin , Cell Signaling Technology , 88145 , Unconjugate.

Techniques: Activation Assay, Marker, Expressing, Quantitative RT-PCR

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Deficiency of Stomach-Type Claudin-18 in Mice Induces Gastric Tumor Formation Independent of H pylori Infection

doi: 10.1016/j.jcmgh.2019.03.003

Figure Lengend Snippet: Antibodies Used in Immunofluorescence Images and FACS Analysis

Article Snippet: Active β-catenin , Cell Signaling Technology , 88145 , Unconjugate.

Techniques: Immunofluorescence