Journal: Toxicological Research

Article Title: Comparison of structural characteristics and molecular markers of rabbit skin, pig skin, and reconstructed human epidermis for an ex vivo human skin model

doi: 10.1007/s43188-023-00185-1

Figure Lengend Snippet: Thickness measurement of each layer of the epidermis in each model. A Immunohistochemistry staining images of human skin, Keraskin, pig skin, and rabbit skin for LOR, KRT10, and KRT5. Scale bars = 30 µm. B Quantification of the thickness of the granular layer stained by LOR. Data are presented as the mean ± SEM. * p < 0.05, **** p < 0.0001; unpaired Student’s t test, n = 20. C Quantification of the thickness of the spinous layer stained by KRT10. Data are presented as the mean ± SEM. ns, not significant ( p > 0.05); ** p < 0.05, **** p < 0.0001; unpaired Student’s t test, n = 20. D Quantification of the thickness of the basal layer stained by KRT5. Data are presented as the mean ± SEM. ns, not significant ( p > 0.05); * p < 0.05, **** p < 0.0001; unpaired Student’s t test, n = 20

Article Snippet: The sections were then incubated with the following primary antibodies overnight at 4 °C: anti-LOR (Abcam, ab24722, 1:2000), anti-KRT10 (Abcam, ab76318, 1:3000), anti-KRT5 (Abcam, ab52635, 1:100,000), anti-MKI67 (Abcam, ab16667, 1:1000), anti-FLG (Novus, NBP1-87,528, 1:1000), anti-CLDN1 (Cell Signaling Technology, 13,255, 1:200), and anti-CDH1 (Cell Signaling Technology, 3195, 1:400).

Techniques: Immunohistochemistry, Staining

Journal: JID Innovations

Article Title: Targeting TGF-β1/miR-21 Pathway in Keratinocytes Reveals Protective Effects of Silymarin on Imiquimod-Induced Psoriasis Mouse Model

doi: 10.1016/j.xjidi.2022.100175

Figure Lengend Snippet: SM blocks the keratinocyte proliferation‒differentiation switch through the inhibition of NOTCH3. ( a ) RT-qPCR quantification of relative expression of K1, K10, IVL, and FLG to GAPDH levels in HaCaT cells treated with 10 ng/ml TGF-β1 in the presence or absence of indicated concentrations of SM for 24 h. ( b‒e ) HaCaT cells were maintained in low calcium (0.07 mM)‒containing medium and then switched to high calcium (1.8 mM)‒containing medium for 9 days in the presence or absence of indicated concentrations of SM. ATRA (1 μM) was used as a negative control. ( b ) Immunofluorescence staining of K10. Bar = 20 μM. ( c ) Western blot analysis of IVL and LOR protein levels. β-Actin was used as a protein loading control. Band intensities were quantitatively analyzed using the ImageJ software. ( d ) The relative gene expression of NOTCH3 to GAPDH was analyzed by RT-qPCR. ( e ) Western blot analysis showing the time course of NOTCH3 protein expression in HaCaT cells treated with SM (62 μg/ml). β-Actin was used as a loading control. Data are presented as mean ± SEM and are representative of three independent experiments performed in triplicate. Two-tailed unpaired t -test was applied for a to test for significance between TGF-β1‒treated cells with SM and TGF-β1‒treated cells alone. ANOVA test was used for d for multiple comparison between the different groups. Two-tailed unpaired t -test was applied for e to test for significance between CaCl2-treated cells with SM and CaCl2-treated cells alone at the 24 h time point, chosen arbitrarily. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗ P < 0.001. ATRA, all-trans retinoid acid; CaCl2, calcium chloride; FDR, false discovery rate; GO, Gene Ontology; h, hour; IVL, involucrin; K, keratin; LOR, loricrin; ns, not significant; SM, silymarin.

Article Snippet: After a blocking step with 5% (w/v) nonfat dry milk for 1 hour at RT, membranes were incubated overnight at 4 °C with antibodies specific to AGO2 (1:1,000, number ab32381, Abcam), AMPKα1/2 (1:500, number ab80039, Abcam), phosphorylated Thr183/172-AMPKα1/2 (1:1,000, number sc-101630, Santa Cruz Biotechnology, Dallas, TX), β-actin (1:5,000, number A2066, Sigma-Aldrich), cyclooxygenase 2 (1:500, number 12282T, Cell Signaling Technology, Danvers, MA), cytokeratin 1 (1:500, number ab93652, Abcam), cytokeratin 10 (1:500, number ab9025, Abcam), GAPDH (1:5,000, number ab9485, Abcam), IVL (1:1,000, number I9018, Sigma Aldrich), LOR (1:500, number ab24722, Abcam), NOTCH3 (1:1,000, number 5276S, Cell Signaling Technology), SMAD2/3 (1:1,000, number 3102S, Cell Signaling Technology), phosphorylated Ser465/467-SMAD2 (1:1,000, number 3108S, Cell Signaling Technology), and SREBP-1 (1:250, number sc-13551, Santa Cruz Biotechnology).

Techniques: Inhibition, Quantitative RT-PCR, Expressing, Negative Control, Immunofluorescence, Staining, Western Blot, Software, Two Tailed Test

Journal: JID Innovations

Article Title: Targeting TGF-β1/miR-21 Pathway in Keratinocytes Reveals Protective Effects of Silymarin on Imiquimod-Induced Psoriasis Mouse Model

doi: 10.1016/j.xjidi.2022.100175

Figure Lengend Snippet: Evaluation of the safety and adverse events of topical application of SM on mice skin. SM (9 mg in acetone) was applied on the dorsal skin of mice (n = 7) for 6 consecutive days. Then mice were photographed on day 7 before collecting skin biopsy samples for histology and gene expression analysis. Control mice received no treatment (n = 7). ( a ) Representative pictures of one representative mouse treated with SM over time. ( b ) Representative H&E-stained tissue sections of the back skin of one representative mouse treated with SM. ( c ) Left panel: representative immunohistochemical detection of Ki-67 on tissue sections of mouse back skin of one representative mouse treated with SM. Right panel: quantification of Ki-67 staining intensity (pixels/μm) in the epidermis of SM-treated mice and control, not treated mice. RT-qPCR quantification of the relative expression of ( d ) epidermal differentiation, ( e ) inflammatory cytokines, as well as ( f ) lipogenic markers of SM-treated mice and control, not treated mice. Bar = 100 μM. Data in c , d , e , and f are the mean ± SEM of seven individual mice per group and are representative of one experiment. Two-tailed unpaired t -test was used to test for significance between SM-treated mice and control, not-treated mice. ∗ P < 0.05 and ∗∗ P < 0.01. IVL, involucrin; K, keratin; LOR, loricrin; ns, not significant; SM, silymarin.

Article Snippet: After a blocking step with 5% (w/v) nonfat dry milk for 1 hour at RT, membranes were incubated overnight at 4 °C with antibodies specific to AGO2 (1:1,000, number ab32381, Abcam), AMPKα1/2 (1:500, number ab80039, Abcam), phosphorylated Thr183/172-AMPKα1/2 (1:1,000, number sc-101630, Santa Cruz Biotechnology, Dallas, TX), β-actin (1:5,000, number A2066, Sigma-Aldrich), cyclooxygenase 2 (1:500, number 12282T, Cell Signaling Technology, Danvers, MA), cytokeratin 1 (1:500, number ab93652, Abcam), cytokeratin 10 (1:500, number ab9025, Abcam), GAPDH (1:5,000, number ab9485, Abcam), IVL (1:1,000, number I9018, Sigma Aldrich), LOR (1:500, number ab24722, Abcam), NOTCH3 (1:1,000, number 5276S, Cell Signaling Technology), SMAD2/3 (1:1,000, number 3102S, Cell Signaling Technology), phosphorylated Ser465/467-SMAD2 (1:1,000, number 3108S, Cell Signaling Technology), and SREBP-1 (1:250, number sc-13551, Santa Cruz Biotechnology).

Techniques: Expressing, Staining, Immunohistochemical staining, Quantitative RT-PCR, Two Tailed Test

Journal: Journal of Immunology Research

Article Title: Immune Imbalance in Nasal Polyps of Caucasian Chronic Rhinosinusitis Patients Is Associated with a Downregulation of E-Selectin

doi: 10.1155/2014/959854

Figure Lengend Snippet: TaqMan Gene expression assays used for qPCR.

Article Snippet: Blots were blocked in 20 mM Tris base and 137 mM sodium chloride (TBS) containing 5% milk powder and 0.1% Tween 20 (TBS-T), for 60 min at room temperature and incubated overnight at 4°C with mouse monoclonal antibody against E-selectin (CD62E, ab24723, Abcam PLC, Cambridge, MA, USA), diluted 1 : 500 in TBS-T containing 3% milk powder.

Techniques: Expressing, Amplification

Journal: Journal of Immunology Research

Article Title: Immune Imbalance in Nasal Polyps of Caucasian Chronic Rhinosinusitis Patients Is Associated with a Downregulation of E-Selectin

doi: 10.1155/2014/959854

Figure Lengend Snippet: Scatter plots of adhesion molecule expression in nasal polyps compared to associated inferior turbinates using microarray and qPCR. Each single dot shows the relative expression of the target molecule in nasal polyps compared to associated inferior turbinates of one patient and median is indicated as horizontal bar . (a) Microarray analysis: nasal polyp tissues showed higher gene expression levels of the adhesion molecules P-selectin (1.62-fold, n = 7), PSGL1 (1.74-fold, n = 7), and VCAM1 (2.16-fold, n = 7) compared to associated inferior turbinates. ICAM1 was unregulated (1.03-fold, n = 7) and E-selectin was strongly downregulated in nasal polyps (0.32-fold, n = 7). (b) qPCR analysis: lower median expression of E-selectin (0.22-fold, n = 14) and higher median expression of P-selectin (2.37-fold, n = 15) were observed in nasal polyps. The median expression of PSGL1, ICAM1, and VCAM1 showed no significant difference between nasal polyps and inferior turbinates: PSGL1 1.24-fold ( n = 10), ICAM1 1.24-fold ( n = 10), and VCAM1 0.85-fold ( n = 10). The mRNA levels of E-selectin in nasal polyps decreased significantly ( P = 0.004) compared to the inferior turbinates and P-selectin increased significantly ( P = 0.0025). PSGL1, ICAM1, and VCAM1 were expressed at similar levels.

Article Snippet: Blots were blocked in 20 mM Tris base and 137 mM sodium chloride (TBS) containing 5% milk powder and 0.1% Tween 20 (TBS-T), for 60 min at room temperature and incubated overnight at 4°C with mouse monoclonal antibody against E-selectin (CD62E, ab24723, Abcam PLC, Cambridge, MA, USA), diluted 1 : 500 in TBS-T containing 3% milk powder.

Techniques: Expressing, Microarray

Journal: Journal of Immunology Research

Article Title: Immune Imbalance in Nasal Polyps of Caucasian Chronic Rhinosinusitis Patients Is Associated with a Downregulation of E-Selectin

doi: 10.1155/2014/959854

Figure Lengend Snippet: Scatter plots of CD31 and adhesion molecule expression in nasal polyps compared to associated inferior turbinates using qPCR. Each single dot shows the relative expression of the target molecule in nasal polyps compared to associated inferior turbinates of one patient and median is indicated as horizontal bar . (a) CD31 is not differentially expressed (0.93-fold, n = 7) between nasal polyps and inferior turbinates; (b) adhesion molecule expression in nasal polyps normalized to CD31. E-selectin (0.22-fold, n = 7), P-selectin (1.82-fold, n = 7), PSGL1 (1.29-fold, n = 7), ICAM1 (0.93-fold, n = 7), and VCAM1 (1.56-fold, n = 7). (c) Adhesion molecule expression in nasal polyps normalized to β -actin. E-selectin (0.31-fold, n = 7), P-selectin (1.97-fold, n = 7), PSGL1 (1.34-fold, n = 7), ICAM1 (1.28-fold, n = 7), and VCAM1 (1.08-fold, n = 7). CD31 normalized data differ insignificantly compared to the β -actin normalized data.

Article Snippet: Blots were blocked in 20 mM Tris base and 137 mM sodium chloride (TBS) containing 5% milk powder and 0.1% Tween 20 (TBS-T), for 60 min at room temperature and incubated overnight at 4°C with mouse monoclonal antibody against E-selectin (CD62E, ab24723, Abcam PLC, Cambridge, MA, USA), diluted 1 : 500 in TBS-T containing 3% milk powder.

Techniques: Expressing

Journal: Journal of Immunology Research

Article Title: Immune Imbalance in Nasal Polyps of Caucasian Chronic Rhinosinusitis Patients Is Associated with a Downregulation of E-Selectin

doi: 10.1155/2014/959854

Figure Lengend Snippet: Comparison of CD31 and β -actin normalized qPCR data.

Article Snippet: Blots were blocked in 20 mM Tris base and 137 mM sodium chloride (TBS) containing 5% milk powder and 0.1% Tween 20 (TBS-T), for 60 min at room temperature and incubated overnight at 4°C with mouse monoclonal antibody against E-selectin (CD62E, ab24723, Abcam PLC, Cambridge, MA, USA), diluted 1 : 500 in TBS-T containing 3% milk powder.

Techniques: Expressing

Journal: Journal of Immunology Research

Article Title: Immune Imbalance in Nasal Polyps of Caucasian Chronic Rhinosinusitis Patients Is Associated with a Downregulation of E-Selectin

doi: 10.1155/2014/959854

Figure Lengend Snippet: Western blotting exhibited lower E-selectin expression in nasal polyps (P) than in the inferior turbinate (IT) of chronic rhinosinusitis patients ( n = 8).

Article Snippet: Blots were blocked in 20 mM Tris base and 137 mM sodium chloride (TBS) containing 5% milk powder and 0.1% Tween 20 (TBS-T), for 60 min at room temperature and incubated overnight at 4°C with mouse monoclonal antibody against E-selectin (CD62E, ab24723, Abcam PLC, Cambridge, MA, USA), diluted 1 : 500 in TBS-T containing 3% milk powder.

Techniques: Western Blot, Expressing

Journal: Journal of Immunology Research

Article Title: Immune Imbalance in Nasal Polyps of Caucasian Chronic Rhinosinusitis Patients Is Associated with a Downregulation of E-Selectin

doi: 10.1155/2014/959854

Figure Lengend Snippet: (a): Expression and localization of E-selectin in inferior turbinates and nasal polyps of chronic rhinosinusitis patients ( n = 10). Immunohistochemical staining was performed on frozen tissue sections of inferior turbinate (left) and nasal polyp (right). We were able to detect E-selectin positive cells in the endothelium of inferior turbinates and nasal polyps. E-selectin was expressed at high level in the inferior turbinates, whereas in nasal polyps E-selectin was infrequently expressed and always at a low level. (b) Expression and localization of P-selectin in inferior turbinates and nasal polyps of chronic rhinosinusitis patients ( n = 10). Immunohistochemical staining was performed on frozen tissue sections of inferior turbinate (left) and nasal polyp (right). P-selectin was detected in the endothelium of inferior turbinates and nasal polyps. P-selectin was always expressed at high levels in both inferior turbinates and nasal polyps.

Article Snippet: Blots were blocked in 20 mM Tris base and 137 mM sodium chloride (TBS) containing 5% milk powder and 0.1% Tween 20 (TBS-T), for 60 min at room temperature and incubated overnight at 4°C with mouse monoclonal antibody against E-selectin (CD62E, ab24723, Abcam PLC, Cambridge, MA, USA), diluted 1 : 500 in TBS-T containing 3% milk powder.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: World Journal of Gastroenterology

Article Title: Radiomics signature: A potential biomarker for β-arrestin1 phosphorylation prediction in hepatocellular carcinoma

doi: 10.3748/wjg.v28.i14.1479

Figure Lengend Snippet: Univariate and multivariate regression analyses of the p-β-arrestin1-positive and p-β-arrestin1-negative groups in the training cohort

Article Snippet: Subsequently, the tissue slides were incubated with primary antibodies using rabbit anti-human p-β-arrestin1 polyclonal antibody (Abcam Biotechnology, ab247229; diluted, 1:200) at 4 °C overnight, followed by incubation with secondary antibodies (cat # K5007; Dako).

Techniques:

Journal: World Journal of Gastroenterology

Article Title: Radiomics signature: A potential biomarker for β-arrestin1 phosphorylation prediction in hepatocellular carcinoma

doi: 10.3748/wjg.v28.i14.1479

Figure Lengend Snippet: Performance of the three models. A: The developed clinico-radiological (CR) nomogram; B: The developed clinico-radiological-radiomics (CRR) nomogram. Predictor points are found on the uppermost point scale that corresponds to each variable. On the bottom scale, the points for all variables are added and translated into a β-arrestin1 phosphorylation positivity probability. C: Comparison of receiver operating characteristic (ROC) curves of the radiomics model, CR model and CRR model in the training cohort; D: Comparison of receiver operating characteristic (ROC) curves of the radiomics model, CR model and CRR model in the validation cohort. E: Calibration curves of the three models in the training cohort; F: Calibration curves of the three models in the validation cohort. The actual high expression of p-β-arrestin1 is represented on the y-axis, and the predicted probability is represented on the x-axis. The closer fit of the solid line to the ideal black dotted line indicates a better calibration.

Article Snippet: Subsequently, the tissue slides were incubated with primary antibodies using rabbit anti-human p-β-arrestin1 polyclonal antibody (Abcam Biotechnology, ab247229; diluted, 1:200) at 4 °C overnight, followed by incubation with secondary antibodies (cat # K5007; Dako).

Techniques: Expressing

Journal: World Journal of Gastroenterology

Article Title: Radiomics signature: A potential biomarker for β-arrestin1 phosphorylation prediction in hepatocellular carcinoma

doi: 10.3748/wjg.v28.i14.1479

Figure Lengend Snippet: Representative images of contrast-enhanced computed tomography and β-Arrestin1 phosphorylation (magnification, × 100). A: CT images of a 45-year-old man with a 6.3-cm hepatocellular carcinoma (HCC) in the right liver lobe in the plain phase; B: The tumor shows heterogeneous hyperenhancement in the arterial phase; C: The tumor shows washout at the portal venous phase with intratumor necrosis, an ill-defined capsule and a non-smooth tumor margin. D: Immunohistochemical staining shows a β-arrestin1 phosphorylation-negative status at 100× magnification.

Article Snippet: Subsequently, the tissue slides were incubated with primary antibodies using rabbit anti-human p-β-arrestin1 polyclonal antibody (Abcam Biotechnology, ab247229; diluted, 1:200) at 4 °C overnight, followed by incubation with secondary antibodies (cat # K5007; Dako).

Techniques: Computed Tomography, Immunohistochemical staining, Staining

Journal: World Journal of Gastroenterology

Article Title: Radiomics signature: A potential biomarker for β-arrestin1 phosphorylation prediction in hepatocellular carcinoma

doi: 10.3748/wjg.v28.i14.1479

Figure Lengend Snippet: Diagnostic performance of the three models for predicting β-arrestin1 phosphorylation-positive hepatocellular carcinoma

Article Snippet: Subsequently, the tissue slides were incubated with primary antibodies using rabbit anti-human p-β-arrestin1 polyclonal antibody (Abcam Biotechnology, ab247229; diluted, 1:200) at 4 °C overnight, followed by incubation with secondary antibodies (cat # K5007; Dako).

Techniques: Diagnostic Assay

Journal: World Journal of Gastroenterology

Article Title: Radiomics signature: A potential biomarker for β-arrestin1 phosphorylation prediction in hepatocellular carcinoma

doi: 10.3748/wjg.v28.i14.1479

Figure Lengend Snippet: Decision curve analysis for each model. A: Decision curve analysis in the training cohort; B: Decision curve analysis in the validation cohort. The y-axis measures the net benefit, and the x-axis is the threshold probability. The gray line represents the hypothesis that all patients are β-arrestin1 phosphorylation-positive. The black line represents the hypothesis that all patients are β-arrestin1 phosphorylation-negative. Among the three models, the clinico-radiological-radiomics (CRR) model provided the highest net benefit compared with the radiomics and clinico-radiological (CR) models.

Article Snippet: Subsequently, the tissue slides were incubated with primary antibodies using rabbit anti-human p-β-arrestin1 polyclonal antibody (Abcam Biotechnology, ab247229; diluted, 1:200) at 4 °C overnight, followed by incubation with secondary antibodies (cat # K5007; Dako).

Techniques:

Journal: World Journal of Gastroenterology

Article Title: Radiomics signature: A potential biomarker for β-arrestin1 phosphorylation prediction in hepatocellular carcinoma

doi: 10.3748/wjg.v28.i14.1479

Figure Lengend Snippet: Overall survival (OS) curve analysis. A: The OS curve estimates by clinic-radiological-radiomics model in patients with β-Arrestin1 phosphorylation positive and β-Arrestin1 phosphorylation negative in the training cohort; B: The OS curve estimates by clinic-radiological-radiomics model in patients with β-Arrestin1 phosphorylation positive and β-Arrestin1 phosphorylation negative in the validation cohort.

Article Snippet: Subsequently, the tissue slides were incubated with primary antibodies using rabbit anti-human p-β-arrestin1 polyclonal antibody (Abcam Biotechnology, ab247229; diluted, 1:200) at 4 °C overnight, followed by incubation with secondary antibodies (cat # K5007; Dako).

Techniques: