Journal: Breast Cancer : Targets and Therapy

Article Title: ALKBH5 Promotes Breast Cancer Stemness Through Regulating Wnt/β-Catenin Signaling

doi: 10.2147/BCTT.S520532

Figure Lengend Snippet: Silencing ALKBH5 inhibits sphere formation capability of MCF-7 cells. ( A ) Mammosphere-forming ability of the stable cell lines was measured in ultra-low attachment plates. ( B ) The number of spheroids formed was quantified. ( C ) Representative images of spheroids formed in ELDA assay. ( D ) ELDA expressing median values from ALKBH5 -knockdown MCF-7 cells (Control, black curves; ALKBH5 -knockdown, red curves). The table shows the stem cell frequency and p value calculated by ELDA. ( E ) The mRNA level of stemness-related genes ( NANOG, EPCAM, CK19 ) was measured by qRT-PCR in ALKBH5 -knockdown MCF-7 spheroids. ( F ) The protein levels of AFP and CK19 in nonsphere cell line. Statistical significance was determined using two-tailed T -test, with p<0.05 considered significant. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Membranes were blocked in 5% non-fat milk and then incubated with primary antibodies targeting ALKBH5 (Novus, NBP1-82188), β-catenin (Abcam, ab32572), AFP (Abcam, ab284388), CK19 (ABclonal, A19040), C-Myc (SANTA CRUZ, sc-40), Cyclin D1 (Cell Signaling Technology, 2922), GAPDH (ABclonal, A19056), N6-methyladenosine (ABclonal, A22411) overnight at 4°C.

Techniques: Stable Transfection, Expressing, Knockdown, Control, Quantitative RT-PCR, Two Tailed Test

Journal: Biomedicines

Article Title: Necroptosis-Related Prognostic Model for Pancreatic Carcinoma Reveals Its Invasion and Metastasis Potential through Hybrid EMT and Immune Escape

doi: 10.3390/biomedicines11061738

Figure Lengend Snippet: Five NRDEGs were identified based on GEO and TCGA database. ( A ) Volcano plot of differentially expressed genes from GSE28735. ( B ) Volcano plot of differentially expressed genes from GSE15471. ( C ) Venn diagram of the intersection of DEGs and necroptosis-related genes. ( D ) Differential expression of 13 NRDEGs in the TCGA database. * means p < 0.05, *** means p < 0.001 and ns means no sense. ( E – I ) Kaplan–Meier curves of NRDEGs: ( E ) KRT7 ( p < 0.001), ( F ) IGF2BP3 ( p = 0.026), ( G ) PKM ( p = 0.007), ( H ) KRT19 ( p = 0.008), ( I ) CXCL5 ( p = 0.047).

Article Snippet: Primary antibodies against KRT7 (1:50, Wuhan Boster Bio-Engineering Limited Company, Wuhan, China), KRT19 (1:50, Wuhan Boster Bio-Engineering Limited Company, Wuhan, China), IGF2BP3 (1:50, Signalway Antibody, MD, USA), and CXCL5 (1:50, Sangon Biotechnology, Shanghai, China) were added to these tissue sections, followed by incubation at 4 °C overnight.

Techniques: Quantitative Proteomics

Journal: Biomedicines

Article Title: Necroptosis-Related Prognostic Model for Pancreatic Carcinoma Reveals Its Invasion and Metastasis Potential through Hybrid EMT and Immune Escape

doi: 10.3390/biomedicines11061738

Figure Lengend Snippet: A necroptosis-related risk prognostic model was successfully constructed. ( A ) Ten-time cross-validation for tuning parameter selection in the LASSO model. ( B ) LASSO coefficient profiles. ( C ) The risk score, survival status, and heat map of model genes in patients with PC. ( D ) Kaplan–Meier curves showed that high-risk group had a worse prognosis than the low-risk group. ( E , F ) PCA and t-SNE visualization of low- and high-risk clusters. ( G ) ROC curves of KRT7, KRT19, CXCL5 and IGF2BP2. ( H ) ROC curves of prognostic model for 1, 3, 5 years, ( I ) ROC curves of risk score and clinicopathological characteristics. ( J , K ) Univariate and multivariate Cox regression analysis of clinicopathological features of PC associated with OS. ( L ) Clinical characteristics and prognostic model were used to establish a predictive nomogram. * means p < 0.05, ** means p < 0.01 ( M ) Heatmap of model genes and clinical parameters.

Article Snippet: Primary antibodies against KRT7 (1:50, Wuhan Boster Bio-Engineering Limited Company, Wuhan, China), KRT19 (1:50, Wuhan Boster Bio-Engineering Limited Company, Wuhan, China), IGF2BP3 (1:50, Signalway Antibody, MD, USA), and CXCL5 (1:50, Sangon Biotechnology, Shanghai, China) were added to these tissue sections, followed by incubation at 4 °C overnight.

Techniques: Construct, Biomarker Discovery, Selection

Journal: Biomedicines

Article Title: Necroptosis-Related Prognostic Model for Pancreatic Carcinoma Reveals Its Invasion and Metastasis Potential through Hybrid EMT and Immune Escape

doi: 10.3390/biomedicines11061738

Figure Lengend Snippet: Model genes were involved in EMT of PC. ( A ) The correlation heat map between model genes and EMT-related genes. ( B ) KRT7 expression in primary as well as metastatic tumors in the EMTome database. ( C ) KRT19 expression in primary as well as metastatic tumors in EMTome database. ( D ) Gene mutations of KRT7 in various cancers in EMTome database. ( E ) Gene mutations of KRT19 in various cancers in EMTome database.

Article Snippet: Primary antibodies against KRT7 (1:50, Wuhan Boster Bio-Engineering Limited Company, Wuhan, China), KRT19 (1:50, Wuhan Boster Bio-Engineering Limited Company, Wuhan, China), IGF2BP3 (1:50, Signalway Antibody, MD, USA), and CXCL5 (1:50, Sangon Biotechnology, Shanghai, China) were added to these tissue sections, followed by incubation at 4 °C overnight.

Techniques: Expressing

Journal: Biomedicines

Article Title: Necroptosis-Related Prognostic Model for Pancreatic Carcinoma Reveals Its Invasion and Metastasis Potential through Hybrid EMT and Immune Escape

doi: 10.3390/biomedicines11061738

Figure Lengend Snippet: Model genes regulated immunity of PC. ( A – D ) The correlation of model genes with various immune cells: ( A ) KRT7, ( B ) KRT19, ( C ) CXCL5, ( D ) IGF2BP3. ( E – H ) Histogram of correlation between model genes and immune cells (Th2, TFH cells). ( I – L ) Scatter plot of correlation between model genes and immune cells (Th2, TFH cells). ( M ) Expressions of immune checkpoints between the high- and low-risk groups. * means p < 0.05, ** means p < 0.01, *** means p < 0.001 and ns means no sense.

Article Snippet: Primary antibodies against KRT7 (1:50, Wuhan Boster Bio-Engineering Limited Company, Wuhan, China), KRT19 (1:50, Wuhan Boster Bio-Engineering Limited Company, Wuhan, China), IGF2BP3 (1:50, Signalway Antibody, MD, USA), and CXCL5 (1:50, Sangon Biotechnology, Shanghai, China) were added to these tissue sections, followed by incubation at 4 °C overnight.

Techniques:

Journal: Biomedicines

Article Title: Necroptosis-Related Prognostic Model for Pancreatic Carcinoma Reveals Its Invasion and Metastasis Potential through Hybrid EMT and Immune Escape

doi: 10.3390/biomedicines11061738

Figure Lengend Snippet: Model genes were upregulated in clinical PC samples and PC cell lines. ( A ) HE, Masson and IHC staining of model genes including KRT7, KRT19, IGF2BP3 and CXCL5 in clinical PC tissues and corresponding adjacent tumor tissues. ( B ) Western blot and histograms of model genes in hTERT-HPNE, PANC-1 and PL45. ( C ) Immunofluorescence (×400) of model genes in hTERT-HPNE, PANC-1 and PL45. hTERT-HPNE is normal human pancreatic ductal cell line, and PANC-1 and PL45 are human pancreatic ductal carcinoma cell lines. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. hTERT-HPNE.

Article Snippet: Primary antibodies against KRT7 (1:50, Wuhan Boster Bio-Engineering Limited Company, Wuhan, China), KRT19 (1:50, Wuhan Boster Bio-Engineering Limited Company, Wuhan, China), IGF2BP3 (1:50, Signalway Antibody, MD, USA), and CXCL5 (1:50, Sangon Biotechnology, Shanghai, China) were added to these tissue sections, followed by incubation at 4 °C overnight.

Techniques: Immunohistochemistry, Western Blot, Immunofluorescence

Journal: Biomedicines

Article Title: Necroptosis-Related Prognostic Model for Pancreatic Carcinoma Reveals Its Invasion and Metastasis Potential through Hybrid EMT and Immune Escape

doi: 10.3390/biomedicines11061738

Figure Lengend Snippet: Prognostic model genes for PC reveals its invasion and metastasis potential through hybrid EMT and immune escape. Hypoxic pancreatic cancer cells are more likely to suffer necroptosis to induce inflammation and immune responses in tumor tissues, which further enhance fibrosis and heterogeneity of tumor, thus the fibrotic and inflammatory circumstance together with immune evasion induced by Th2 assist PC cells to escape from immune elimination. Furthermore, our model genes promote tumor invasion and metastasis by promoting EMT especially hybrid EMT of PC cells that KRT7 and KRT19 are involved in EMT as epithelial components, while CXCL5 and IGF2BP3 act as regulatory factors to regulate EMT. Ultimately, the above effects together promote tumor invasion and metastasis.

Article Snippet: Primary antibodies against KRT7 (1:50, Wuhan Boster Bio-Engineering Limited Company, Wuhan, China), KRT19 (1:50, Wuhan Boster Bio-Engineering Limited Company, Wuhan, China), IGF2BP3 (1:50, Signalway Antibody, MD, USA), and CXCL5 (1:50, Sangon Biotechnology, Shanghai, China) were added to these tissue sections, followed by incubation at 4 °C overnight.

Techniques:

Journal: American Journal of Translational Research

Article Title: Effects of human amnion-derived mesenchymal stem cells and conditioned medium in rats with sclerosing cholangitis

doi:

Figure Lengend Snippet: Primer sequences for quantitative reverse transcription-polymerase chain reaction

Article Snippet: Tissue sections were stained with anti-rat cytokeratin 19 (CK19) antibody (dilution 1:100; Proteintech, Chicago, IL, USA) to assess biliary hyperplasia.

Techniques: Sequencing

Journal: American Journal of Translational Research

Article Title: Effects of human amnion-derived mesenchymal stem cells and conditioned medium in rats with sclerosing cholangitis

doi:

Figure Lengend Snippet: Effects of human amnion-derived mesenchymal stem cells (hAMSCs) and conditioned medium (CM) obtained from hAMSCs on biliary hyperplasia. A. Cytokeratin 19 (CK19) expression. B. Quantitative reverse transcription-polymerase chain reaction for CK19. The stained areas were measured from 10 sections per sample in each low-power field. Scale bars, 100 μm. Values are expressed as the mean ± SEM (N = 6-10 animals/group). *P < 0.05 and **P < 0.01 vs. the control group. †P < 0.05 vs. the alpha-naphthylisothiocyanate (ANIT) group.

Article Snippet: Tissue sections were stained with anti-rat cytokeratin 19 (CK19) antibody (dilution 1:100; Proteintech, Chicago, IL, USA) to assess biliary hyperplasia.

Techniques: Derivative Assay, Expressing, Reverse Transcription, Polymerase Chain Reaction, Staining, Control

Journal: Heliyon

Article Title: AHR regulates liver enlargement and regeneration through the YAP signaling pathway

doi: 10.1016/j.heliyon.2024.e37265

Figure Lengend Snippet: Activation of AHR promoted liver enlargement and regeneration in wild-type mice. (A) Wild-type mice were treated with vehicle or YH439 (100 mg/kg/d) for 10 days. (B) The relative liver/body weight ratios (n = 5). (C) Representative liver photos of vehicle or YH439-treated mice. (D) CTNNB1 staining, H&E staining were performed to measure the cell size around CV area. Quantification of cell size (n = 5). (E)CK19/Ki67 staining was performed to measure the cell proliferation around PV area. Quantification the number of Ki67 + cells (n = 5). Data are expressed as means ± SD; *p < 0.05, **p < 0.01 and ***p < 0.001; Student's t-test. Scale bar = 50 μm.

Article Snippet: For immunohistochemical staining, Anti-Ki67 (1:100 dilution, BOSTER, Cat# M00254-8, RRID: AB_3081764 ), Anti-CTNNB1(1:100 dilution BOSTER, Cat#PA1212, RRID: AB_3082601 ), Anti-CK19(1:100 dilution BOSTER, Cat#PB9715, RRID: AB_3082234 )/Anti- Ki67 (1:100 dilution), CTNNB1(1:100 dilution)/Ki67(1:100 dilution), Anti-Sox9(1:100 dilution BOSTER, Cat#BM4268, RRID: AB_2941988 )/Anti- Ki67 (1:100 dilution), Anti-Sox9(1:100 dilution)/Anti- Ki67 (1:100 dilution), Anti-Sox9(1:100 dilution)/Anti-HNF4α (1:100 dilution, Santa Cat#sc-374229, RRID: AB_10989766 ) were used as primary antibody.

Techniques: Activation Assay, Staining

Journal: Heliyon

Article Title: AHR regulates liver enlargement and regeneration through the YAP signaling pathway

doi: 10.1016/j.heliyon.2024.e37265

Figure Lengend Snippet: AHR activation-induced liver enlargement and regeneration is abolished in Ahr ΔHep mice. (A) Ahr fl/fl.ALB-CreERT2 and Ahr ΔHep mice were treated with vehicle or YH439 (100 mg/kg/d) for 10 days. (B) The relative liver/body weight ratios (n = 5). (C) Representative liver photos of vehicle or YH439-treated mice. (D) CTNNB1 staining, H&E staining were performed to measure the cell size around CV area. Quantification of cell size (n = 5). (E)CK19/Ki67 staining was performed to measure the cell proliferation around PV area. Quantification the number of Ki67 + cells (n = 5). Data are expressed as means ± SD; *p < 0.05, **p < 0.01 and ***p < 0.001; Student's t-test. Scale bar = 50 μm.

Article Snippet: For immunohistochemical staining, Anti-Ki67 (1:100 dilution, BOSTER, Cat# M00254-8, RRID: AB_3081764 ), Anti-CTNNB1(1:100 dilution BOSTER, Cat#PA1212, RRID: AB_3082601 ), Anti-CK19(1:100 dilution BOSTER, Cat#PB9715, RRID: AB_3082234 )/Anti- Ki67 (1:100 dilution), CTNNB1(1:100 dilution)/Ki67(1:100 dilution), Anti-Sox9(1:100 dilution BOSTER, Cat#BM4268, RRID: AB_2941988 )/Anti- Ki67 (1:100 dilution), Anti-Sox9(1:100 dilution)/Anti- Ki67 (1:100 dilution), Anti-Sox9(1:100 dilution)/Anti-HNF4α (1:100 dilution, Santa Cat#sc-374229, RRID: AB_10989766 ) were used as primary antibody.

Techniques: Activation Assay, Staining

Journal: Heliyon

Article Title: AHR regulates liver enlargement and regeneration through the YAP signaling pathway

doi: 10.1016/j.heliyon.2024.e37265

Figure Lengend Snippet: AHR activation promotes liver regeneration (A) Ahr fl/fl.ALB-CreERT2 and Ahr ΔHep mice were treated with vehicle or YH439 (100 mg/kg/d) for 2 days following PHx. (B) (B) CTNNB1 staining, H&E staining were performed to measure the cell size around CV area. Quantification of cell size (n = 5). Scale bar = 50 μm. (C)CK19/Ki67 staining was performed to measure the cell proliferation around PV area. Quantification the number of Ki67 + cells (n = 5). Scale bar = 20 μm. Data are expressed as means ± SD; *p < 0.05 and **p < 0.01; Student's t-test.

Article Snippet: For immunohistochemical staining, Anti-Ki67 (1:100 dilution, BOSTER, Cat# M00254-8, RRID: AB_3081764 ), Anti-CTNNB1(1:100 dilution BOSTER, Cat#PA1212, RRID: AB_3082601 ), Anti-CK19(1:100 dilution BOSTER, Cat#PB9715, RRID: AB_3082234 )/Anti- Ki67 (1:100 dilution), CTNNB1(1:100 dilution)/Ki67(1:100 dilution), Anti-Sox9(1:100 dilution BOSTER, Cat#BM4268, RRID: AB_2941988 )/Anti- Ki67 (1:100 dilution), Anti-Sox9(1:100 dilution)/Anti- Ki67 (1:100 dilution), Anti-Sox9(1:100 dilution)/Anti-HNF4α (1:100 dilution, Santa Cat#sc-374229, RRID: AB_10989766 ) were used as primary antibody.

Techniques: Activation Assay, Staining

Journal: Heliyon

Article Title: AHR regulates liver enlargement and regeneration through the YAP signaling pathway

doi: 10.1016/j.heliyon.2024.e37265

Figure Lengend Snippet: YAP-TEAD interaction is involved in AHR-induced liver enlargement and regeneration. (A) Corn oil-treated mice and verteporfin-treated mice were administered vehicle or YH439 (100 mg/kg/d) for 5 days. (B) The relative liver/body weight ratios (n = 5). (C) Representative liver photos of vehicle or YH439-treated mice. (D) CTNNB1 staining, H&E staining were performed to measure the cell size around CV area. Quantification of cell size (n = 5). (E)CK19/Ki67 staining were performed to measure the cell proliferation around PV area. Quantification the number of Ki67 + cells (n = 5). Data are expressed as means ± SD; *p < 0.05, **p < 0.01 and ***p < 0.001; Student's t-test. Scale bar = 50 μm.

Article Snippet: For immunohistochemical staining, Anti-Ki67 (1:100 dilution, BOSTER, Cat# M00254-8, RRID: AB_3081764 ), Anti-CTNNB1(1:100 dilution BOSTER, Cat#PA1212, RRID: AB_3082601 ), Anti-CK19(1:100 dilution BOSTER, Cat#PB9715, RRID: AB_3082234 )/Anti- Ki67 (1:100 dilution), CTNNB1(1:100 dilution)/Ki67(1:100 dilution), Anti-Sox9(1:100 dilution BOSTER, Cat#BM4268, RRID: AB_2941988 )/Anti- Ki67 (1:100 dilution), Anti-Sox9(1:100 dilution)/Anti- Ki67 (1:100 dilution), Anti-Sox9(1:100 dilution)/Anti-HNF4α (1:100 dilution, Santa Cat#sc-374229, RRID: AB_10989766 ) were used as primary antibody.

Techniques: Staining

Journal: Heliyon

Article Title: AHR regulates liver enlargement and regeneration through the YAP signaling pathway

doi: 10.1016/j.heliyon.2024.e37265

Figure Lengend Snippet: Yap is involved in AHR-induced liver enlargement and regeneration. (A) Yap fl/fl.ALB-CreERT2 and Yap ΔHep were treated with vehicle or YH439 (100 mg/kg/d) for 10 days. (B) The relative liver/body weight ratios (n = 5). (C) Representative liver photos of vehicle or YH439-treated mice. (D) CTNNB1 staining, H&E staining were performed to measure the cell size around CV area. Quantification of cell size (n = 5). (E)CK19/Ki67 staining was performed to measure the cell proliferation around PV area. Quantification the number of Ki67 + cells (n = 5). Data are expressed as means ± SD; *p < 0.05, **p < 0.01 and ***p < 0.001; Student's t-test. Scale bar = 50 μm.

Article Snippet: For immunohistochemical staining, Anti-Ki67 (1:100 dilution, BOSTER, Cat# M00254-8, RRID: AB_3081764 ), Anti-CTNNB1(1:100 dilution BOSTER, Cat#PA1212, RRID: AB_3082601 ), Anti-CK19(1:100 dilution BOSTER, Cat#PB9715, RRID: AB_3082234 )/Anti- Ki67 (1:100 dilution), CTNNB1(1:100 dilution)/Ki67(1:100 dilution), Anti-Sox9(1:100 dilution BOSTER, Cat#BM4268, RRID: AB_2941988 )/Anti- Ki67 (1:100 dilution), Anti-Sox9(1:100 dilution)/Anti- Ki67 (1:100 dilution), Anti-Sox9(1:100 dilution)/Anti-HNF4α (1:100 dilution, Santa Cat#sc-374229, RRID: AB_10989766 ) were used as primary antibody.

Techniques: Staining

Journal: Stem Cell Research & Therapy

Article Title: Heme oxygenase-1-modified bone marrow mesenchymal stem cells combined with normothermic machine perfusion to protect donation after circulatory death liver grafts

doi: 10.1186/s13287-020-01736-1

Figure Lengend Snippet: Expression of CK19 in biliary epithelial cells of the transplanted liver. A Immunofluorescence staining for CK19 (× 200, red tag) showed that the percentage of CK19-positive cells among biliary epithelial cells in the HBP group was significantly higher than that in the other groups. B Expression of CK19 in the transplanted liver analyzed by western blotting. C Relative expression of the CK19 protein (CK19/β-actin) showed that the relative expression of CK19 in the HBP group was significantly higher than that in the other groups

Article Snippet: Samples were dehydrated, embedded in paraffin, sectioned, and subjected to HO-1 and cytokeratin 19 (CK19) staining (Proteintech, Wuhan, China).

Techniques: Expressing, Immunofluorescence, Staining, Western Blot