Journal: Journal of Oncology

Article Title: A New Model for Predicting Nonsentinel Lymph Node Metastasis in Early-Stage Breast Cancer Using MMP15

doi: 10.1155/2022/8675705

Figure Lengend Snippet: The expression of matrix metalloproteinase 15 (MMP15) in the cytoplasm of positive sentinel lymph nodes (SLNs); the cells were magnified 400 times. (a) The expression of MMP15 was positive. (b) The expression of MMP15 was negative.

Article Snippet: All sections were incubated with one or two drops of primary MMP15 antibody (rabbit polyclonal antibody, Abcam, ab15475, 1/100) for 60 min at room temperature.

Techniques: Expressing

Journal: Journal of Oncology

Article Title: A New Model for Predicting Nonsentinel Lymph Node Metastasis in Early-Stage Breast Cancer Using MMP15

doi: 10.1155/2022/8675705

Figure Lengend Snippet: Comparison of clinicopathological characteristics of the training cohort and validation cohort.

Article Snippet: All sections were incubated with one or two drops of primary MMP15 antibody (rabbit polyclonal antibody, Abcam, ab15475, 1/100) for 60 min at room temperature.

Techniques: Expressing, Over Expression, Standard Deviation

Journal: Journal of Oncology

Article Title: A New Model for Predicting Nonsentinel Lymph Node Metastasis in Early-Stage Breast Cancer Using MMP15

doi: 10.1155/2022/8675705

Figure Lengend Snippet: Univariate logistic regression analysis of factors related to NSLN metastases.

Article Snippet: All sections were incubated with one or two drops of primary MMP15 antibody (rabbit polyclonal antibody, Abcam, ab15475, 1/100) for 60 min at room temperature.

Techniques: Expressing, Over Expression

Journal: Journal of Oncology

Article Title: A New Model for Predicting Nonsentinel Lymph Node Metastasis in Early-Stage Breast Cancer Using MMP15

doi: 10.1155/2022/8675705

Figure Lengend Snippet: Multivariate logistic regression analysis of factors related to NSLN metastases.

Article Snippet: All sections were incubated with one or two drops of primary MMP15 antibody (rabbit polyclonal antibody, Abcam, ab15475, 1/100) for 60 min at room temperature.

Techniques: Expressing

Journal: Journal of Oncology

Article Title: A New Model for Predicting Nonsentinel Lymph Node Metastasis in Early-Stage Breast Cancer Using MMP15

doi: 10.1155/2022/8675705

Figure Lengend Snippet: Nomogram to predict the probability of nonsentinel lymph node (NSLN) metastasis in sentinel lymph nodes (SLNs)-positive breast cancer patients. The new model included five variables: matrix metalloproteinase 15 (MMP15) expression in the cytoplasm, number of positive SLNs, neural invasion, tumor size, and lymphovascular invasion. After each variable was assigned, it corresponded to the score of the first row in the nomogram and then the five scores were summed to get the total score. From this score, the NSLN metastasis probability could be obtained.

Article Snippet: All sections were incubated with one or two drops of primary MMP15 antibody (rabbit polyclonal antibody, Abcam, ab15475, 1/100) for 60 min at room temperature.

Techniques: Expressing

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Figure Lengend Snippet: SYCP3 (red) and SYCP1 (green). (A,A') Zygotene trivalents (T1 and T2) and their schematic representation. The distal ends of the acrocentric chromosomes are synapsed with the metacentric chromosomes (arrowheads) whereas the proximal ends are unsynapsed (arrows). Two unsynapsed proximal ends are engaged in a heterologous end-to-end connection (asterisk). (B,B') Early pachytene. Trivalents show co-localization of SYCP3 and SYCP1 in the synapsed regions (arrowheads) whereas unsynapsed regions (arrows) that do not present SYCP1. One trivalent is associated with the X chromosome by a short segment of SC (red arrowhead). The single Y chromosome appears self-synapsed. (C,C') Complete homologous synapsis of two acrocentric chromosomes with the metacentric chromosome. (D,D') Heterologous synapsis between two acrocentric chromosomes involving a small chromosome region (arrowhead). (E,E') Partial heterologous synapsis of the acrocentric chromosomes involving a long chromosome region (arrowhead). Notice the unsynapsed chromosomal segment in the heterologous region (arrows). (F,F') Open trivalent. The proximal region of one acrocentric and the interstitial region of the metacentric chromosome do not present SYCP1, while the other acrocentric chromosome presents SYCP1 in its unsynapsed segment. (G,G') During diplotene, the heterologous synapsis between acrocentric chromosomes persists (arrowheads) even when desynapsis begins in the trivalent.

Article Snippet: The slides were placed in PBS and incubated with the following primary antibodies: mouse anti-SYCP3 1∶100 (Abcam, Ab12452); rabbit anti-SYCP3 1∶100 (Abcam, Ab15093); rabbit anti-SYCP1 1∶100 (Abcam, Ab15087); rabbit anti RAD51 1∶50 (Calbiochem, PC130); mouse anti-phospho-histone H2AX (Ser139) 1∶1000, clone JBW301 (Upstate, 05–636); goat anti ATR 1∶80 (Santa Cruz Biotechnology, sc-1887); mouse anti ubiquityl-histone H2A 1∶15, clone E6C5 (Upstate, 05–678); mouse anti GMP-1 (SUMO-1) 1∶50 (Zymed, 33–2400); mouse RIK2D3 1∶100 that recognizes the XMR protein in the testis , kindly provided by Denise Escalier (Université Paris 5, Paris, France).

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Figure Lengend Snippet: γH2AX appears at zygotene distributed in the whole chromatin while ATR appears as discrete foci scattered along the AEs/LEs as revealed by SYCP3 protein. At the beginning of pachytene, γH2AX becomes restricted to the chromatin surrounding unsynapsed AEs. Accumulation of SUMO 1, monoubiquitinated H2A, and XMR on unsynapsed chromatin occurs through the early pachytene stage, before the accumulation of ATR in the unsynapsed chromatin at mid-pachytene. These proteins remain associated to the unsynapsed chromatin until late diplotene, indicating that spermatocytes with unsynapsed trivalents skip pachytene arrest and progress further into later stages of meiosis. Newly desynapsed chromosomal regions at diplotene do not incorporate any of the MSUC-related proteins.

Article Snippet: The slides were placed in PBS and incubated with the following primary antibodies: mouse anti-SYCP3 1∶100 (Abcam, Ab12452); rabbit anti-SYCP3 1∶100 (Abcam, Ab15093); rabbit anti-SYCP1 1∶100 (Abcam, Ab15087); rabbit anti RAD51 1∶50 (Calbiochem, PC130); mouse anti-phospho-histone H2AX (Ser139) 1∶1000, clone JBW301 (Upstate, 05–636); goat anti ATR 1∶80 (Santa Cruz Biotechnology, sc-1887); mouse anti ubiquityl-histone H2A 1∶15, clone E6C5 (Upstate, 05–678); mouse anti GMP-1 (SUMO-1) 1∶50 (Zymed, 33–2400); mouse RIK2D3 1∶100 that recognizes the XMR protein in the testis , kindly provided by Denise Escalier (Université Paris 5, Paris, France).

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Figure Lengend Snippet: The expression of human interferon gamma (IFNγ) by different coat protein (CP)-deficient Potexvirus-based vectors in N. benthamiana. ( A ). Schematic representation of Potexvirus-based expression cassettes, in which the CP coding regions of Potato virus X (PVX), Bamboo mosaic virus (BaMV), and Foxtail mosaic virus (FoMV) were replaced with that of the target protein (TP), IFNγ. These constructs were named pKPIFNγ (a), pKBIFNγ (b), and pKFIFNγ (c), with the expected molecular weight of the respective TP indicated in the adjacent table (d). ( B ). Analysis of TP expression in inoculated leaves. Total protein extracts, corresponding to 1 mg fresh weight in 1 mL of extraction buffer, were prepared from infiltrated leaf tissue at 3, 5, and 7 days-post infiltration (DPI) and analyzed by SDS-PAGE, followed by staining with CBS. IB analysis of proteins transferred to the membrane used anti-IFNγ as the primary antibody and goat-anti-rabbit IgG horseradish peroxidase conjugate as the secondary antibody. M, Marker; H, Healthy leaf sample; P, Positive control, purified mIFNγ protein derived from E. coli (100 ng in CBS and 2.5 ng in IB). The positions of various TP species and reference proteins were indicated on the right of the panel. The mono- and dimeric forms of IFNγ were indicated by M or D, respectively. ( C ). Quantification by ELISA. The ELISA quantification shows the IFNγ yield relative to fresh weight of leaves (μg/g). Statistical analysis was performed using ANOVA. p -value of <0.001 was considered significant. The double 35S promoters were indicated by the two thick arrows. The nos terminator is represented by the black box at the end of the construct.

Article Snippet: To verify that the higher signal intensity for mIFNγ was not resulted from the preferences of the primary antibody, we have performed immunoblot analyses using different primary antibodies with different specificity, including anti-IFNγ (ab133566 from abcam Co., specific to N-terminal 1–100 amino acids of native IFNγ), anti-His tag (GTX115045 from GeneTex Co., specific to the 6X His tag), and the original anti-mIFNγ ( ).

Techniques: Expressing, Construct, Molecular Weight, SDS Page, Staining, Marker, Positive Control, Purification, Derivative Assay, Enzyme-linked Immunosorbent Assay

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Figure Lengend Snippet: Codon usage optimization of IFNγ or mIFNγ gene based on the codon bias of N. benthamiana . ( A ). Schematic representation of Bamboo mosaic virus (BaMV)-based expression cassettes, in which BaMV CP coding region was replaced with TP (IFNγ or mIFNγ, for full-length or mature protein without signal peptide, respectively) using restriction sites Mlu I and Spe I. The coding sequence of either IFNγ or mIFNγ were further optimized according to the codon bias for N. benthamiana species to generate IFNγ-1/-2 and mIFNγ-1/-2, respectively. ( B ). Analysis of TP expression in inoculated leaves. Total protein extracts were prepared from infiltrated leaf tissue at 3 and 5 DPI and analyzed by SDS-PAGE, followed by staining with CBS and IB analysis with anti-mIFNγ as primary antibody and goat-anti-rabbit IgG alkaline phosphatase conjugate as secondary antibody. M, Marker; H, Healthy leaf; P, Positive control, purified mIFNγ protein derived from E. coli (250 ng for CBS and 25 ng for IB). ( C ). Quantification with densitometer. Densitometric quantification showing the percentage values of M or D TP relative to RuBisCO Small subunit, rbcS. Statistical analysis was performed using ANOVA. p -value of <0.001 was considered significant. ( D ). Northern blot analysis of wild-type or chimeric BaMV RNA in infiltrated leaves at 3 and 5 DPI. BaMV genomic RNA, and the subgenomic RNAs for triple gene block proteins (TGPsgRNA) and CP (CPsgRNA) were detected with a BaMV-specific probe, respectively (vector, pKB△C His as control). The bottom panel shows the amount of rRNA in each sample, stained with ethidium bromide as the loading control.

Article Snippet: To verify that the higher signal intensity for mIFNγ was not resulted from the preferences of the primary antibody, we have performed immunoblot analyses using different primary antibodies with different specificity, including anti-IFNγ (ab133566 from abcam Co., specific to N-terminal 1–100 amino acids of native IFNγ), anti-His tag (GTX115045 from GeneTex Co., specific to the 6X His tag), and the original anti-mIFNγ ( ).

Techniques: Expressing, Sequencing, SDS Page, Staining, Marker, Positive Control, Purification, Derivative Assay, Northern Blot, Blocking Assay, Plasmid Preparation

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Figure Lengend Snippet: Details of antibodies used in the study.

Article Snippet: Anti-mouse/rat/human Glut1 , EPR3915 , Alexa Fluor 647 , 1/100 , Rabbit , abcam , ab195020 , AB_2783877.

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