Journal: American Journal of Cancer Research

Article Title: MEX3A suppresses proliferation and EMT via inhibiting Akt signaling pathway in cervical cancer

doi:

Figure Lengend Snippet: The expression level of MEX3A protein in tissues and its functions on viability. A. Representative MEX3A staining figure (high or low expression) in tissue microarray. Scale bars: 50 μm. B. The correlation between MEX3A expression with clinical stage and lymph node involvement. C. MEX3A expression levels in cervical cancer cells and normal cervical epithelial cells at transcriptional and translational level. D. The efficacy of MEX3A knockdown in C33A and MEX3A overexpression in SiHa were measured by Western blotting.

Article Snippet: Cell culture and reagents The human cervical cancer cells C33A, SiHa, Caski and MS751 cells, as well as normal cervical cancer cells H8 were obtained from American Type Culture Collection (ATCC, USA).

Techniques: Expressing, Staining, Microarray, Knockdown, Over Expression, Western Blot

Journal: American Journal of Cancer Research

Article Title: MEX3A suppresses proliferation and EMT via inhibiting Akt signaling pathway in cervical cancer

doi:

Figure Lengend Snippet: MEX3A inhibits the viability and induced cell cycle arrest. A. MEX3A knockdown promoted viability of C33A cells, while MEX3A overexpression suppressed viability of SiHa cells by CCK-8 assay. *P < 0.05, **P < 0.01, ***P < 0.001. B. Left panel: MEX3A knockdown promoted proliferation of C33A cells, and MEX3A overexpression suppressed proliferation of SiHa cells by BrdU assay. Scale bar: 50 μm. Right panel: Quantitative results were also performed. C. MEX3A knockdown decreased the percentage of G0/G1 phase but increased cells at S phase in C33A cells, while MEX3A overexpression showed the opposite results in SiHa cells by detection of flow cytometry.

Article Snippet: Cell culture and reagents The human cervical cancer cells C33A, SiHa, Caski and MS751 cells, as well as normal cervical cancer cells H8 were obtained from American Type Culture Collection (ATCC, USA).

Techniques: Knockdown, Over Expression, CCK-8 Assay, BrdU Staining, Flow Cytometry

Journal: American Journal of Cancer Research

Article Title: MEX3A suppresses proliferation and EMT via inhibiting Akt signaling pathway in cervical cancer

doi:

Figure Lengend Snippet: MEX3A inhibits the migration and invasion of cervical cancer cells. A. Left panel: Cell migration ability was detected by wound healing assay in C33A and SiHa cells after transfection with MEX3A shRNA or MEX3A cDNA. Right panel: Quantitative data was shown for left panel. B. Transwell assays were performed in cells with MEX3A downregulation or upregulation. Right panel: Quantitative analysis for left panel. **P < 0.01, ***P < 0.001.

Article Snippet: Cell culture and reagents The human cervical cancer cells C33A, SiHa, Caski and MS751 cells, as well as normal cervical cancer cells H8 were obtained from American Type Culture Collection (ATCC, USA).

Techniques: Migration, Wound Healing Assay, Transfection, shRNA

Journal: American Journal of Cancer Research

Article Title: MEX3A suppresses proliferation and EMT via inhibiting Akt signaling pathway in cervical cancer

doi:

Figure Lengend Snippet: MEX3A suppresses tumor growth in vivo. (A) The picture of xenografts was obtained at 35 day after injection of transfected C33A and SiHa cells in nude mice. (B) Tumors were collected and each tumor weight was measured. (C) The tumor volume was assessed and calculated according to the formula (L × W2)/2. The data were dispalyed as mean ± SD. (D, E) The representative pictures for H&E staining (top) and immunohistochemical staining of E-cadherin, β-catenin of C33A (D) and SiHa (E) subcutaneous tumor samples from the different groups. Scale bar: 50 μm. ***P < 0.001.

Article Snippet: Cell culture and reagents The human cervical cancer cells C33A, SiHa, Caski and MS751 cells, as well as normal cervical cancer cells H8 were obtained from American Type Culture Collection (ATCC, USA).

Techniques: In Vivo, Injection, Transfection, Staining, Immunohistochemical staining

Journal: iScience

Article Title: Autophagy ablation in skeletal muscles worsens sepsis-induced muscle wasting, impairs whole-body metabolism, and decreases survival

doi: 10.1016/j.isci.2023.107475

Figure Lengend Snippet:

Article Snippet: mouse IgG2b monoclonal anti-MHC type I , DSHB , Cat# BA-F8; RRID: AB_10572253.

Techniques: Western Blot, Immunofluorescence, Muscles, Recombinant, Microscopy, SYBR Green Assay, Reverse Transcription, Microarray, Gene Expression, Software, Control, Irradiation, Staining

Journal: International Journal of Molecular Sciences

Article Title: Transcriptional Analysis of Hair Follicle-Derived Keratinocytes from Donors with Atopic Dermatitis Reveals Enhanced Induction of IL32 Gene by IFN-γ

doi: 10.3390/ijms14023215

Figure Lengend Snippet: Genes showing differential expression between FDKs from donors with atopic dermatitis (AD) and from non-atopic controls (Non-AD) by microarray analysis, as determined by comparison analysis using a permutation test. The log 2 ratio (the Cy5 to Cy3 fluorescence intensity using the same Cy3-labelled probe, consisting of a mixture of NHEK extracts and Universal Human Reference RNA, as a reference) was used as a expression level. For comparative studies of gene expression between AD-FDKs and Non-AD-FDKs, a permutation test was performed (number of permutations: 500) and 827 genes were extracted. In this figure, 11 genes that exhibit a difference of at least 2-fold and meet the criterion p < 0.05 are visualized as the tree after hierarchical clustering (Distance; pearson uncentered, Linkage; complete).

Article Snippet: Probes with poor-quality signals (signal/noise < 1.2) and those with null data at a frequency of more than 35% in all samples were filtered out using the microarray data analysis software Expressionist (GeneData AG, Basel, Switzerland); after this procedure, 13,722 probes remained.

Techniques: Quantitative Proteomics, Microarray, Comparison, Fluorescence, Expressing, Gene Expression

Journal:

Article Title: Pathogenic Natural Antibodies Recognizing Annexin IV Are Required to Develop Intestinal Ischemia-Reperfusion Injury 1

doi: 10.4049/jimmunol.0803980

Figure Lengend Snippet: Characterization of mAb B4 epitope expression. A, Representative flow cytometric histograms show binding of mAb B4 to a single cell suspension of IEC (top), splenocytes (middle) and thymocytes (bottom). Cells were incubated with mAb B4, and bound Ab was detected by using anti-mouse IgM (μ-chain specific) for IEC and thymocytes. In the case of splenocytes, mAb B4 labeled with biotin was used. B and C, Monoclonal Ab B4 epitope expression was determined by Western blot analysis of isolated from thymus, spleen or intestine cell in (B) or lysates were prepared from whole organs in (C). Data are representative of two independent experiments. D, Binding of mAb B4 to proteins typically found as targets of polyreactive natural Abs was studied by micro-array analysis. The positive control for mAb B4 binding are C1q and anti-mouse IgM antibody, positive controls for the micro-array are mAb NC-17D8 and polyclonal IgM, and the negative control is designated blank.

Article Snippet: Polyclonal rabbit anti-annexin IV Ab was obtained from ProteinTech Group, and anti-6xHis mAb was obtained from Novagen.

Techniques: Expressing, Binding Assay, Suspension, Incubation, Labeling, Western Blot, Isolation, Microarray, Positive Control, Negative Control

Journal:

Article Title: Pathogenic Natural Antibodies Recognizing Annexin IV Are Required to Develop Intestinal Ischemia-Reperfusion Injury 1

doi: 10.4049/jimmunol.0803980

Figure Lengend Snippet: Identification of the antigen recognized by mAb B4. A, Lysates prepared from isolated IEC were separated using a sequential three gel separation protocol as described in Materials and Methods. The presence of the B4 antigen was confirmed by Western blot analysis (right). Two gel samples (gray color boxes) were analyzed by MS. B, The MASCOT search results for the protein isolated from first sample was identified as mouse annexin IV based on the highest score number of 785; 78 matched peptides covering 53% of the annexin IV sequence were identified (underlying bold letters). C, Lysates of untransformed F-293 cells (lane 1) and F-293 cells transformed with the pSecTag2/Hygro B expression vector carrying annexin IV insert (F-293-A4) (lane 3) together with culture supernatants from transformed cells (lane 2) were probed by Western blot analysis with mAb B4. D, Flow cytometric analysis of F-293 cells expressing recombinant annexin IV. F-293-A4 cells were probed in flow cytometry with mAb B4. E, Supernatant (lane1) and lysate (lane 2) from F-293-A4 cells expressing recombinant annexin IV that had been incubated in PBS alone, or supernatant (lane 3) and lysate (lane 4) from the same cells incubated with 0.5 M EDTA, were probed by Western blot analysis with mAb B4. Recombinant annexin IV was not released from F-293-A4 cells in the absence of free Ca2+.

Article Snippet: Polyclonal rabbit anti-annexin IV Ab was obtained from ProteinTech Group, and anti-6xHis mAb was obtained from Novagen.

Techniques: Isolation, Western Blot, Sequencing, Transformation Assay, Expressing, Plasmid Preparation, Recombinant, Flow Cytometry, Incubation

Journal:

Article Title: Pathogenic Natural Antibodies Recognizing Annexin IV Are Required to Develop Intestinal Ischemia-Reperfusion Injury 1

doi: 10.4049/jimmunol.0803980

Figure Lengend Snippet: A, Intestinal IR injury is ameliorated in wildtype C57Bl/6 mice receiving recombinant annexin IV prior to the ischemic phase. Reduction of the of IR induced injury to the level of sham operated animals (B6 sham) was observed in wild type mice when they were injected with 50 μg/mouse of annexin IV 5 min prior to the reperfusion phase (A4 IR). Injury in mice pre-injected with control CPK19 protein (CP K19 IR) was comparable with the injury in C57Bl/6 mice undergoing IR (B6 IR) (a). Small intestine tissue samples were processed as in Fig. 6 for myeloperoxidase activity (b) and the eicosanoids LTB4 (c) and PGE2 (d). Each bar is the average ± SEM with 3–6 - animals per group. Statistical significance was determined using one way ANOVA. B, Small intestine of annexin IV pre-injected mice (far right), similarly to sham operated wild type mice (far left) does not show C3 deposition in IR induced injury, in contrast to wild type mice (second from left) and mice pre-injected with the CPK19 control protein undergoing IR (second from right).

Article Snippet: Polyclonal rabbit anti-annexin IV Ab was obtained from ProteinTech Group, and anti-6xHis mAb was obtained from Novagen.

Techniques: Recombinant, Injection, Control, Activity Assay

Journal:

Article Title: Pathogenic Natural Antibodies Recognizing Annexin IV Are Required to Develop Intestinal Ischemia-Reperfusion Injury 1

doi: 10.4049/jimmunol.0803980

Figure Lengend Snippet: Presence of natural antibodies to annexin IV. A, Cr2+/+ in contrast to Cr2−/− mice demonstrate higher levels of IgM natural Ab to annexin IV. Serum samples from three cohorts of mice, Cr2−/− (n=9), Cr2+/+ mice (n=14) and negative control Rag1−/− (n=5) mice were evaluated for the presence of natural IgM (left) and IgG (right) Abs to bacterial annexin IV. The figures are representative of two independent experiments. Statistical significance was determined using a Wilcoxson test. B, 9 serum samples from healthy humans and three from agammaglobulinemic patients were tested in the anti-annexin IV ELISA. Substantial levels of anti-annexin IV IgM antibody IgM is present in the sera of normal humans.

Article Snippet: Polyclonal rabbit anti-annexin IV Ab was obtained from ProteinTech Group, and anti-6xHis mAb was obtained from Novagen.

Techniques: Negative Control, Enzyme-linked Immunosorbent Assay

Journal: Oncotarget

Article Title: Dual inhibition of HDAC and EGFR signaling with CUDC-101 induces potent suppression of tumor growth and metastasis in anaplastic thyroid cancer.

doi: 10.18632/oncotarget.3268

Figure Lengend Snippet: Figure 3: CUDC-101 inhibits ATC cell proliferation, and induces cell cycle arrest and apoptosis. (A) Basal expression of HDAC1, HDAC2 and EGFR in ATC cell lines. (B) Cell proliferation assay. Error bars are mean ± SD. (C) Cell cycle analysis after 24 hours of treatment. (D) The Caspase-Glo 3/7 assay after 48 hours of treatment with CUDC-101. *p < 0.05, **p < 0.01, ***p < 0.001. NS, no significant difference.

Article Snippet: Phospho-kinase and apoptosis protein arrays Human phospho-kinase (Catalog # ARY003B) array (detecting site-specific phosphorylation of 43 kinases and 2 related total proteins, and apoptosis array (Catalog # ARY009 detecting the expression level of 35 apoptosisrelated proteins) were purchased from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Proliferation Assay, Cell Cycle Assay, Caspase-Glo Assay

Journal: Oncotarget

Article Title: Dual inhibition of HDAC and EGFR signaling with CUDC-101 induces potent suppression of tumor growth and metastasis in anaplastic thyroid cancer.

doi: 10.18632/oncotarget.3268

Figure Lengend Snippet: Figure 5: Protein targets of CUDC-101 in ATC cells. (A) CUDC-101 inhibits HDACs and MAPK in ATC cells. (B) Targets of CUDC-101 identified by phospho-kinase array. Phospho-kinase arrays were performed using cell lysates treated with and without CUDC-101. Shown are those proteins that were altered with CUDC-101 treatment in both ATC cell lines with > 1.5-fold difference. (C) Differentially expressed apoptotic proteins with and without CUDC-101 treatment. Apoptosis arrays were performed. Shown are those proteins that were altered with CUDC-101 treatment in both ATC cell lines with > 1.5-fold difference. For A–C, ATC cells were treated with the vehicle or CUDC-101 at 1.1 μM for 24 hours. (D) CUDC-101 reduces the expression of survivin, XIAP, and β-catenin. Cells were treated with the vehicle or CUDC-101 for 24 hours.

Article Snippet: Phospho-kinase and apoptosis protein arrays Human phospho-kinase (Catalog # ARY003B) array (detecting site-specific phosphorylation of 43 kinases and 2 related total proteins, and apoptosis array (Catalog # ARY009 detecting the expression level of 35 apoptosisrelated proteins) were purchased from R&D Systems (Minneapolis, MN).

Techniques: Expressing