Journal: Frontiers in Genetics

Article Title: CircARVCF Contributes to Cisplatin Resistance in Gastric Cancer by Altering miR-1205 and FGFR1

doi: 10.3389/fgene.2021.767590

Figure Lengend Snippet: Primers sequences used for qRT-PCR.

Article Snippet: Next, the slides were incubated with anti-FGFR1 (bs-0230R; Bioss) overnight and secondary antibody (bs-0293G-HRP) for 1 h. After that, the signal was developed with DAB solution and counterstained with hematoxylin (Sigma-Aldrich) as previously described ( ).

Techniques:

Journal: Frontiers in Genetics

Article Title: CircARVCF Contributes to Cisplatin Resistance in Gastric Cancer by Altering miR-1205 and FGFR1

doi: 10.3389/fgene.2021.767590

Figure Lengend Snippet: CircARVCF regulated FGFR1 expression by targeting miR-1205. (A) The complementary sequences between FGFR1 and miR-1205 were exhibited. (B,C) The relationship between FGFR1 and miR-1205 was analyzed by dual-luciferase reporter assay. (D,E) The mRNA and protein levels of FGFR1 in GC tissues and normal tissues were measured by qRT-PCR or western blot assay. (F) The protein level of FGFR1 in GES-1, MKN-45 and AGS cells was measured via western blot assay. (G,H) The mRNA and protein levels of FGFR1 in DDP-resistant and DDP-sensitive GC tissues were quantified by qRT-PCR or western blot. (I) The protein level of FGFR1 in MKN-45, MKN-45/DDP, AGS and AGS/DDP cells was measured via western blot assay. (J) The protein level of FGFR1 in MKN-45/DDP and AGS/DDP cells transfected with miR-NC, miR-1205, miR-1205 + pcDNA or miR-1205 + FGFR1 was measured via western blot assay. (K) The protein level of FGFR1 in MKN-45/DDP and AGS/DDP cells transfected with si-NC, si-circARVCF#1, si-circARVCF#1+in-miR-NC or si-circARVCF#1+in-miE-1205 was measured through western blot assay. * p < 0.05.

Article Snippet: Next, the slides were incubated with anti-FGFR1 (bs-0230R; Bioss) overnight and secondary antibody (bs-0293G-HRP) for 1 h. After that, the signal was developed with DAB solution and counterstained with hematoxylin (Sigma-Aldrich) as previously described ( ).

Techniques: Expressing, Luciferase, Reporter Assay, Quantitative RT-PCR, Western Blot, Transfection

Journal: Frontiers in Genetics

Article Title: CircARVCF Contributes to Cisplatin Resistance in Gastric Cancer by Altering miR-1205 and FGFR1

doi: 10.3389/fgene.2021.767590

Figure Lengend Snippet: MiR-1205 regulated the DDP resistance and malignant behaviors of DDP-resistant GC cells by targeting FGFR1. MKN-45/DDP and AGS/DDP cells were transfected with miR-NC, miR-1205, miR-1205 + pcDNA or miR-1205 + FGFR1. (A,B) DDP resistance was analyzed by CCK-8 assay. (C–F) The colony formation, migration, invasion and apoptosis of MKN-45/DDP and AGS/DDP cells were evaluated by colony formation assay, transwell assay and flow cytometry analysis, respectively. (G,H) The protein levels of MRP1, MDR1, Bcl-2 and Bax in MKN-45/DDP and AGS/DDP cells were measured via western blot assay. * p < 0.05.

Article Snippet: Next, the slides were incubated with anti-FGFR1 (bs-0230R; Bioss) overnight and secondary antibody (bs-0293G-HRP) for 1 h. After that, the signal was developed with DAB solution and counterstained with hematoxylin (Sigma-Aldrich) as previously described ( ).

Techniques: Transfection, CCK-8 Assay, Migration, Colony Assay, Transwell Assay, Flow Cytometry, Western Blot

Journal: Frontiers in Genetics

Article Title: CircARVCF Contributes to Cisplatin Resistance in Gastric Cancer by Altering miR-1205 and FGFR1

doi: 10.3389/fgene.2021.767590

Figure Lengend Snippet: CircARVCF enhanced DDP resistance in GC in vivo . (A,B) Tumor volume and tumor weight were examined. (C,D) The levels of circARVCF and miR-1205 in the tumors were examined by qRT-PCR. (E,F) The level of FGFR1 in the tumors was examined by western blot assay and IHC assay. * p < 0.05.

Article Snippet: Next, the slides were incubated with anti-FGFR1 (bs-0230R; Bioss) overnight and secondary antibody (bs-0293G-HRP) for 1 h. After that, the signal was developed with DAB solution and counterstained with hematoxylin (Sigma-Aldrich) as previously described ( ).

Techniques: In Vivo, Quantitative RT-PCR, Western Blot

Journal: Frontiers in Genetics

Article Title: CircARVCF Contributes to Cisplatin Resistance in Gastric Cancer by Altering miR-1205 and FGFR1

doi: 10.3389/fgene.2021.767590

Figure Lengend Snippet: The frame diagram of circARVCF/miR-1205/FGFR1 axis in regulating DDP resistance and cell progression in GC.

Article Snippet: Next, the slides were incubated with anti-FGFR1 (bs-0230R; Bioss) overnight and secondary antibody (bs-0293G-HRP) for 1 h. After that, the signal was developed with DAB solution and counterstained with hematoxylin (Sigma-Aldrich) as previously described ( ).

Techniques:

Journal: Nature cancer

Article Title: Molecular analysis of primary melanoma T cells identifies patients at risk for metastatic recurrence

doi: 10.1038/s43018-019-0019-5

Figure Lengend Snippet: Reassessment by multiplex immunohistochemistry to determine the relative abundance of CD3, CD8, CD4, FoxP3, CD39 and CD103. (a) Representative merged image of CD3, CD4 and CD8 staining; n=57 primary melanoma samples. (b) The amount of CD8+, CD4+FoxP3- and CD4+FoxP3+ cells is variable in both TCFr high as well as TCFr low samples independent of their recurrence status as illustrated by the stacked bar plots. (c) Representative merged image of CD8, CD39 and CD103 staining, n=57 primary melanoma samples. (d) Putative tumor specific CD8+ T cells (CD8+CD39+ and CD8+CD39+CD103+) vary in frequency in both non-recurring and recurring TCFr high and TCFr low samples. (e) TCFr by HTS does not correlate with the number of CD8+, CD4+, Tregs and tumor specific CD8+ T cells (CD8+CD39+ and CD8+CD39+CD103+) in the T cell infiltrate. Line=regression line, grey shading = 95% confidence interval, Spearman’s correlation test; n=57 primary melanoma samples. (f) Grouped boxplots of CD8+, CD4+, Tregs, CD8+CD39+ and CD8+CD39+CD103+ cells as percentage of all T cells in the infiltrate colorized by a 20% TCFr cut-off (≥ 20% TCFr in green, < 20% TCFr in orange) and recurrence status. There is no difference in abundance of CD8+ (Kruskal-Wallis Test pKW = 0.263), CD4+ (pKW = 0.263), CD8+CD39+ (pKW = 0.077) nor CD8+CD39+CD103+ (pKW = 0.500) between TCFr high and low samples. Tregs differed (pKW = 0.018) between the four groups since TCFr high non-recurring samples had fewer detectable CD4+FoxP3+ cells than TCFr low recurring samples two-sided Wilcoxon Rank Sum Test with post-hoc Benjamini-Hochberg correction, p = 0.012). Group sizes of primary melanoma samples: TCFr high non-rec, n=16, TCFr high rec, n=14; TCFr low non-rec, n=12; TCFr low rec, n=15. Box plots: the bold line indicates the median; box illustrates lower and upper quartiles; whiskers show the lowest and highest data point still within 1.5x of interquartile range from lower or upper quartile, dots are outliers.

Article Snippet: Then, slides were incubated with the primary antibody for FoxP3 (Abcam, 236A1E7, 1:1000), CD103 (Abcam, EPR4166(2), 1:800), CD8 (Dako, C8/144B, 1:1000), CD4 (Biocare Medical, 4B12, 1:400), CD3 (Cell Marque, MRQ-39, 1:2000) or CD39 (Abcam, {"type":"entrez-protein","attrs":{"text":"EPR20627","term_id":"523387756","term_text":"EPR20627"}} EPR20627 , 1:2000), made up in either Da Vinci Green (Biocare Medical), Antibody Diluent/Block (Perkin Elmer) or Van Gough Yellow (Biocare Medical), for 30 minutes.

Techniques: Multiplex Assay, Immunohistochemistry, Staining

Journal: Oncotarget

Article Title: Downregulation of BRCA1-BRCA2-containing complex subunit 3 sensitizes glioma cells to temozolomide

doi:

Figure Lengend Snippet: Human brain tissue slide used for this study contained 24 cases of patients with different grades of gliomas in duplicates. The tissue slide was subjected to immunohistochemistry staining using anti-BRCC3 antibody (Abcam). The representative images show BRCC3 immunoreactivity in normal human cortical tissue (A) grade I astrocytoma (B) grade II astrocytoma (C) grade III anaplastic astrocytoma (D) grade IV glioblastoma multiforme (E) . Experiments were repeated using anti-BRCC3 antibody from ProSci with similar observations. The staining was photographed under microscope with four images taken from each case. BRCC3 immunoreactivity of normal brain tissue and different grades of glioma were evaluated using ImageJ software (F) . Cells with BRCC3 immunostaining were selected through threshold setting of ImageJ software. The data are referred as immunoreactivity score (IRS) representing the average intensity of BRCC3-positive cells normalized over the intensity of background. ** p < 0.01, versus normal tissue. Scale bar in A-E, 100 μm.

Article Snippet: Human brain tumor tissue slide (Pantomics Inc., Richmond, CA, USA) was first heated at 60 ° C for 30 min before deparaffinization using xylene.

Techniques: Immunohistochemistry, Staining, Microscopy, Software, Immunostaining

Journal: BMC Cardiovascular Disorders

Article Title: Circular RNA expression profile and its potential regulative role in human abdominal aortic aneurysm

doi: 10.1186/s12872-020-01374-8

Figure Lengend Snippet: circRNAs expression profiles detected by microarray in the AAA group and control group. a The box plot shows the nearly identical distributions of normalized intensity values from the aortic samples of the AAA and control group. b The scatter plot is built to assess the expression variation of circRNAs between the two groups. The X and Y axes indicate the normalized intensity values of each circRNAs from the AAA and control group. The dots above the upper green line and below the lower green line represent the dysregulated circRNAs with a fold change (FC) > 2.0 between the two groups. c The volcano plot presents differentially expressed circRNAs in AAA. The vertical lines correspond to 2-fold upregulation and downregulation, the horizontal line indicates P value of 0.05. The red dots represent the differentially expressed circRNAs (FC > 2.0 and P value < 0.05). d Hierarchical clustering analysis reveals a distinguishable expression profile of circRNAs between the AAA and control group. Each column indicates an aortic sample, each row represents a circRNA. The red and green color indicate high and low expression level, respectively. e Chromosomal distribution of the differentially expressed circRNAs between the two groups

Article Snippet: The fragmented labeled cRNAs were hybridized onto the circRNA expression microarray slide (Arraystar Human circRNA Array V2).

Techniques: Expressing, Microarray, Control

Journal: BMC Cardiovascular Disorders

Article Title: Circular RNA expression profile and its potential regulative role in human abdominal aortic aneurysm

doi: 10.1186/s12872-020-01374-8

Figure Lengend Snippet: Validation of six randomly selected dysregulated circRNAs by qRT-PCR. Each circRNA was evaluated at least three times and compared with the results of microarray. The Y axis indicates the fold change of AAA vs control of each circRNA

Article Snippet: The fragmented labeled cRNAs were hybridized onto the circRNA expression microarray slide (Arraystar Human circRNA Array V2).

Techniques: Biomarker Discovery, Quantitative RT-PCR, Microarray, Control

Journal: BMC Cardiovascular Disorders

Article Title: Circular RNA expression profile and its potential regulative role in human abdominal aortic aneurysm

doi: 10.1186/s12872-020-01374-8

Figure Lengend Snippet: The predicted circRNA/miRNA interaction networks for six randomly selected circRNAs. a , b The red nodes indicate upregulated circRNAs. c - f The blue nodes represent downregulated circRNAs. The green nodes are five complementary binding miRNAs of each circRNA

Article Snippet: The fragmented labeled cRNAs were hybridized onto the circRNA expression microarray slide (Arraystar Human circRNA Array V2).

Techniques: Binding Assay