Journal: Oncology Reports

Article Title: 5-Aminolevurinic acid inhibits the proliferation of bladder cancer cells by activating heme synthesis

doi: 10.3892/or.2022.8401

Figure Lengend Snippet: Expression of ferritin, proliferating cell nuclear antigen, survivin, and cytokeratin 20 in bladder tumors in the control group (images on the left) and 5-ALA group (images in the middle) and in a bladder without bladder cancer in the 5-ALA group (images on the right). PCNA, proliferating cell nuclear antigen; CK20, cytokeratin 20; 5-ALA, 5-aminolevulinic acid; Tis, tumor in situ.

Article Snippet: The slides were incubated overnight at 4°C with rabbit monoclonal antibodies against ferritin (product code ab75972, dilution 1:50; Abcam); rabbit polyclonal antibodies against cytokeratin 20 (CK20) (cat. no. bs-1588R; dilution 1:100; Bioss Antibodies); rabbit polyclonal antibodies against survivin (cat. no. 10508-1-AP; dilution 1:500; ProteinTech Group, Inc.), which is specific to the G2/M phase in cells; or rabbit monoclonal antibodies against proliferating cell nuclear antigen (PCNA) (product code ab92552; dilution 1:100; Abcam), which is specific to the S phase in cells.

Techniques: Expressing, Control, In Situ

Journal: Journal of Cellular and Molecular Medicine

Article Title: miR‐126‐5p enhances radiosensitivity of lung adenocarcinoma cells by inhibiting EZH2 via the KLF2/BIRC axis

doi: 10.1111/jcmm.17135

Figure Lengend Snippet: KLF2 affects BIRC5 expression by binding to the promoter region of BIRC5. (A) Correlation analysis of KLF2 and BIRC5 in lung adenocarcinoma. (B) Box plot of BIRC5 expression in lung adenocarcinoma tissue samples. (C) BIRC5 expression in lung adenocarcinoma tissues and adjacent normal tissues determined by RT‐qPCR ( n = 78). (D) BIRC5 protein level in lung adenocarcinoma tissue and adjacent normal tissues determined by immunohistochemical staining ( n = 78, Scale bar = 100 μm). (E) BIRC5 expression in five lung adenocarcinoma cell lines and human embryonic lung fibroblast cell lines determined by RT‐qPCR. (F) The possible binding site of KLF2 and BIRC5 promoter predicted by JASPAR website. (G) The degree of KLF2 enrichment in BIRC5 gene promoter region detected by ChIP‐PCR. (H) The binding site of KLF2 and BIRC5 promoter region verified using the dual‐luciferase reporter gene assay. A549 and H1650 cells were transduced with oe‐KLF2 after irradiation. (I) The transfection efficiency of KLF2 overexpression in A549 and H1650 cells detected by RT‐qPCR. (J) Protein levels of KLF2 and BIRC5 in A549 and H1650 cells measured by Western blot analysis. Data are shown as the mean ± standard deviation of three technical replicates. Data comparisons between two groups were analysed by unpaired t ‐test. Data comparisons among multiple groups were analysed by the one‐way ANOVA with Tukey's post hoc test. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: Afterwards, the samples were blocked with 5% normal goat serum for 15 min and incubated with the following antibodies, rabbit polyclonal antibody against EZH2 (ab186006, 1: 100, Abcam), rabbit polyclonal antibody against KLF2 (PA5‐40591, 1: 100, Invitrogen), or rabbit polyclonal antibody against BIRC5 (10508–1‐AP, 1: 200, Proteintech Group Inc.) at 4°C overnight.

Techniques: Expressing, Binding Assay, Quantitative RT-PCR, Immunohistochemical staining, Staining, Luciferase, Reporter Gene Assay, Transduction, Irradiation, Transfection, Over Expression, Western Blot, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: miR‐126‐5p enhances radiosensitivity of lung adenocarcinoma cells by inhibiting EZH2 via the KLF2/BIRC axis

doi: 10.1111/jcmm.17135

Figure Lengend Snippet: miR‐126‐5p affects the radiosensitivity of A549 cells via the EZH2/KLF2/BIRC5 axis. A549 cells were treated with X‐rays and transduced with miR‐126‐5p mimic alone or combined with si‐KLF2. (A) The transfection efficiency of si‐KLF2 in A549 cells detected by RT‐qPCR. (B) miR‐126‐5p expression in A549 cells determined by RT‐qPCR. (C) Protein levels of EZH2, KLF2 and BIRC5 in A549 cells measured by Western blot analysis. (D) A549 cell apoptosis detected by flow cytometry. (E) The number of γ‐H2AX focus in A549 cells detected by immunofluorescence staining. (F) A549 cell migration detected by Transwell assay. Data are shown as the mean ± standard deviation of three technical replicates. Data comparisons between two groups were analysed by unpaired t ‐test. Data comparisons among multiple groups were analysed by the one‐way ANOVA with Tukey's post hoc test. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: Afterwards, the samples were blocked with 5% normal goat serum for 15 min and incubated with the following antibodies, rabbit polyclonal antibody against EZH2 (ab186006, 1: 100, Abcam), rabbit polyclonal antibody against KLF2 (PA5‐40591, 1: 100, Invitrogen), or rabbit polyclonal antibody against BIRC5 (10508–1‐AP, 1: 200, Proteintech Group Inc.) at 4°C overnight.

Techniques: Transduction, Transfection, Quantitative RT-PCR, Expressing, Western Blot, Flow Cytometry, Immunofluorescence, Staining, Migration, Transwell Assay, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: miR‐126‐5p enhances radiosensitivity of lung adenocarcinoma cells by inhibiting EZH2 via the KLF2/BIRC axis

doi: 10.1111/jcmm.17135

Figure Lengend Snippet: miR‐126‐5p affects the radiosensitivity of lung adenocarcinoma cells via the EZH2/KLF2/BIRC5 axis in vivo. Nude mice were treated with 20 Gy X‐ray alone or combined with lv‐miR‐126‐5p. (A) Tumour volume of nude mice. (B) Xenograft tumour in nude mice. (C) Tumour weight of nude mice. (D) miR‐126‐5p expression in tumour tissues of nude mice determined by RT‐qPCR. (E) Protein levels of EZH2, KLF2, and BIRC5 in tumour tissues of nude mice measured by Western blot analysis. (F) Molecular mechanism of miR‐126‐5p involved in regulating the EZH2/KLF2/BIRC5 axis to improve the radiosensitivity of lung adenocarcinoma cells. Data are shown as the mean ± standard deviation of three technical replicates. Data comparisons between two groups were analysed by unpaired t ‐test. Data comparisons among multiple groups were analysed by the one‐way ANOVA with Tukey's post hoc test. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: Afterwards, the samples were blocked with 5% normal goat serum for 15 min and incubated with the following antibodies, rabbit polyclonal antibody against EZH2 (ab186006, 1: 100, Abcam), rabbit polyclonal antibody against KLF2 (PA5‐40591, 1: 100, Invitrogen), or rabbit polyclonal antibody against BIRC5 (10508–1‐AP, 1: 200, Proteintech Group Inc.) at 4°C overnight.

Techniques: In Vivo, Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation

Journal: Scientific Reports

Article Title: In vivo analysis of onset and progression of retinal degeneration in the Nr2e3 rd7/rd7 mouse model of enhanced S-cone sensitivity syndrome

doi: 10.1038/s41598-021-98271-7

Figure Lengend Snippet: Cell death mechanisms in C57BL/6J Nr2e3 rd7/rd7 retinas. Immunohistochemical analysis of cell death markers on 5 μM-paraffin sections of 1-year-old (1y) C57BL/6J Nr2e3 rd7/rd7 (rd7) ( A , C , E , G , I , K , M ) and wild-type C57BL/6J (BL6) ( B , D , F , H , J , L , N ) retinas. Sections were probed with antibodies raised against Bax ( A , B ), cleaved Caspase-3 ( C , D ), cleaved PARP-1 ( E , F ), MLKL ( G , H ), PAR ( I , J ), Calpain-2 ( K , L ) and Survivin ( M , N ). Bax, cleaved Caspase-3, MLKL and Survivin were detected with a secondary antibody conjugated to Alexa Fluor 594 (red), cleaved PARP-1 and Calpain-2 with a secondary antibody conjugated to Cy5 (red) and PAR with a secondary antibody conjugated to FITC (green). Nuclei were stained with DAPI (blue), namely the outer nuclear layer (ONL) and the inner nuclear layer (INL). Scale bar: 50 µm. ( O ) Qualitative Western blot analysis on six pooled retinas of 21-day (P21) and 1-year-old (1y) C57BL/6J (BL6) and C57BL/6J Nr2e3 rd7/rd7 (rd7) mice. Expression of the 16-kDa survivin and 49-kDa α-tubulin proteins were assessed.

Article Snippet: Membranes were blocked in 5% non-fat dried milk before being immunoassayed using rabbit polyclonal antibodies against survivin (diluted 1/1000; Thermo Fisher PA1-16836), S-opsin and M-opsin (diluted 1/1000; Merck Millipore) and a mouse monoclonal antibody against α-tubulin (1/5000; Sigma, Buchs, Switzerland).

Techniques: Immunohistochemical staining, Staining, Western Blot, Expressing

Journal: Translational Neuroscience

Article Title: Canstatin represses glioma growth by inhibiting formation of VM-like structures

doi: 10.1515/tnsci-2020-0176

Figure Lengend Snippet: Canstatin overexpression suppressed the activation of Akt and decreased the expression of survivin in vitro. (a) Western blotting analysis of total Akt (Akt), phosphorylated Akt (p-Akt), survivin, and HIF-1α protein expression in U87 cells transduced with either lentiviral pCDH vector or pCDH-Canstatin. GAPDH was used as a loading control. (b–e) Relative quantification of Akt, p-Akt, survivin, and HIF-1α protein expression was determined. (f and g) ELISA analysis of MMP-2 and MMP-9 secretion in control and U87 cells overexpressing canstatin. Values are represented as means ± SD, n = 3 each group, * p < 0.05, ** p < 0.01, and *** p < 0.005.

Article Snippet: The primary antibody against survivin rabbit polyclonal antibody (#GB11177) was purchased from Servicebio Inc. (Wuhan, China).

Techniques: Over Expression, Activation Assay, Expressing, In Vitro, Western Blot, Transduction, Plasmid Preparation, Control, Quantitative Proteomics, Enzyme-linked Immunosorbent Assay

Journal: Cancer Science

Article Title: Immunohistological analysis of pancreatic carcinoma after vaccination with survivin 2B peptide: Analysis of an autopsy series

doi: 10.1111/cas.14099

Figure Lengend Snippet: Immunohistochemistry for pancreatic carcinoma. A, HLA class I molecules in the surface of tumor cell. B, Survivin in the nucleus and/or cytoplasm of tumor cell. C, Programmed cell death‐1 ( PD ‐1) in the cell boundary of lymphocytes (arrows). D, Forkhead box P3 ( FOXP 3) in the nucleus of lymphocytes (arrows). E, PD ligand 1 ( PD ‐L1) in the surface of tumor cell. Bar = 50 μm. Original magnification, ×200

Article Snippet: Rabbit polyclonal Ab against survivin (2 μg/mL; Novus Biologicals, Littleton, CO, USA) was used.

Techniques: Immunohistochemistry

Journal: Cancer Science

Article Title: Immunohistological analysis of pancreatic carcinoma after vaccination with survivin 2B peptide: Analysis of an autopsy series

doi: 10.1111/cas.14099

Figure Lengend Snippet: Vaccination schedule for each patient included in the clinical trial who was subsequently investigated at autopsy and the distinctive infiltration pattern of CD 8 + cells in pancreatic carcinoma. A, In Step 1, the patients were treated until diagnosed as having progressive disease by RECIST or clinically apparent progressive disease. In Step 2, all patients who had given their consent were treated with survivin 2B peptide ( SVN ‐2B) and β‐interferon ( IFN β) until diagnosed as immune‐related response criteria (ir RC ) immune‐related progression of disease (ir PD ) or for a maximum 7 mo. B,C, Double staining pictures showing pan‐cytokeratin (red) expressed in tumor cells and CD 8 (brown) in lymphocytes. CD 8 + lymphocyte‐rich (B) and CD 8 + lymphocyte‐poor (C) tumor tissue specimens are shown. There is a small number of infiltrated CD 8 + lymphocytes in the CD 8 + lymphocyte‐poor lesion (C) identified as the internal positive control (arrows). Bar = 50 μm. Original magnification, ×200. D, Heterogenous distribution of CD 8 + lymphocytes in tumor tissue. Dotted line indicates the boundary between the tumor and interstitial tissue. Bar = 100 μm. Original magnification, ×100. E, Schematic diagram of lymphocyte infiltration into tumor tissue composed of tumor cells and interstitial tissue

Article Snippet: Rabbit polyclonal Ab against survivin (2 μg/mL; Novus Biologicals, Littleton, CO, USA) was used.

Techniques: Double Staining, Positive Control

Journal: Cancer Science

Article Title: Immunohistological analysis of pancreatic carcinoma after vaccination with survivin 2B peptide: Analysis of an autopsy series

doi: 10.1111/cas.14099

Figure Lengend Snippet: In situ investigation of infiltration of CD 8 + and programmed cell death‐1 ( PD ‐1) + lymphocytes into pancreatic carcinoma lesions. A,B, Comparison of infiltration of CD 8 + cells in the tumor (A) and interstitial tissue (B) between patients who had and had not received the survivin 2B peptide ( SVN ‐2B) vaccine. C, Number of CD 8 + lymphocytes that infiltrated each tumor lesion. D,E, Comparison of infiltration of PD ‐1 + cells into the tumor (D) and interstitial tissue (E) between patients who had and had not received the SVN ‐2B vaccine. F, Number of PD ‐1 + lymphocytes that infiltrated each lesion. G,H, Correlation between proportions of the PD ‐L1 + tumor and CD 8 + (G) or PD ‐1 + (H) lymphocyte infiltration in the lesions. PD ‐L1 expression on the surface of tumor cells was deemed to be positive. Signal intensity was not evaluated

Article Snippet: Rabbit polyclonal Ab against survivin (2 μg/mL; Novus Biologicals, Littleton, CO, USA) was used.

Techniques: In Situ, Comparison, Expressing

Journal: Cancer Science

Article Title: Immunohistological analysis of pancreatic carcinoma after vaccination with survivin 2B peptide: Analysis of an autopsy series

doi: 10.1111/cas.14099

Figure Lengend Snippet: Immunological effects in peripheral blood and histopathological analysis of pancreatic carcinoma tumor specimens from cases 2 and 7. A,E, Flow cytometry analysis of peripheral blood for CD 8 + T lymphocytes specific for survivin 2B peptide ( SVN ‐2B) peptide on the HLA ‐A24 complex before (left panel) and 8 weeks after (right panel) the first vaccination in case 2 (A) and case 7 (E). B,F, γ‐Interferon ( IFN γ) enzyme‐linked immunospot assay of PBMC s at 8 weeks after the first vaccination for no peptide (left), HIV (center), or SVN ‐2B (right) specific peptide in case 2 (B) and case 7 (F). C,G, Gross appearance of the cut surface of a liver lesion after formalin fixation in case 2 and before fixation in case 7 (G). Arrows indicate metastatic lesions. D,H, Morphological image (H&E stain; left panel) and immunohistochemistry for CD 8 (center panel) and programmed cell death ligand 1 ( PD ‐L1) (right panel) of a metastatic liver lesion in case 2 (D) and case 7 (H). Insets show high‐power magnification

Article Snippet: Rabbit polyclonal Ab against survivin (2 μg/mL; Novus Biologicals, Littleton, CO, USA) was used.

Techniques: Flow Cytometry, Enzyme-linked Immunospot, Staining, Immunohistochemistry

Journal: Oncotarget

Article Title: ASIC1 promotes differentiation of neuroblastoma by negatively regulating Notch signaling pathway

doi: 10.18632/oncotarget.14164

Figure Lengend Snippet: The shASIC1a and pEGFP-ASIC1a constructs were transfected into NS20Y cells and the changes in expression levels of Notch1signaling were then determined by Western blot. A. The levels of endogenous expression of Notch1 and its downstream targeting gene, Survivin were detected in NS20Y cells. Both Notch1and Survivin are constitutively expressed in NS20Y (lane 1 and 5 in upper panel), knock-down of shASIC1a resulted in upregulation of Notch1 gene and its downstream target Survivin gene (upper left panel, lane 1 vs. 2 and lane 3 vs.4); B. While overexpression of ASIC1a downregulated Notch1 and Survivin genes (upper right panel).

Article Snippet: Polyclonal rabbit antibody against Survivin was purchased from Cell Signaling Technology, Inc (Danvers, MA).

Techniques: Construct, Transfection, Expressing, Western Blot, Knockdown, Over Expression

Journal: Oncotarget

Article Title: ASIC1 promotes differentiation of neuroblastoma by negatively regulating Notch signaling pathway

doi: 10.18632/oncotarget.14164

Figure Lengend Snippet: DAPT effectively inhibited the Notch response by causing a reduction on the level of Notch1 and its responsive gene Survivin (lane 1,2,3,4 vs.5,6,7,8 respectively).

Article Snippet: Polyclonal rabbit antibody against Survivin was purchased from Cell Signaling Technology, Inc (Danvers, MA).

Techniques:

Journal: Oncotarget

Article Title: Inhibition of skeletal growth of human prostate cancer by the combination of docetaxel and BKM1644: an aminobisphosphonate derivative

doi: 10.18632/oncotarget.8481

Figure Lengend Snippet: ( A ) BKM1644 (5 μM) inhibits survivin proteins and the phosphorylation of Stat3 at Ser727 and Tyr705 residues in Western blot analysis. ( B ) Top panel: human survivin promoter contains two putative Stat3 binding elements; Bottom panel: BKM1644 selectively inhibits the luciferase activity of pSurvivin-Luc1430 in a dose-dependent manner. The reporters were transfected into C4-2 cells for 24 h prior to further treatment with BKM1644 for 48 h. ( C ) Top panel: Docetaxel treatment (2.5 nM) increases survivin protein expression in a time-dependent manner in C4-2 cells; Bottom panel: BKM1644 (5 μM) antagonizes survivin induction by docetaxel (2 nM) and activates apoptosis in C4-2 cells during a 24-h culture. ( D ) BKM1644 sensitizes C4-2 cells to docetaxel treatment in a dose-dependent manner (72 h).

Article Snippet: Expression of survivin and Ki67 in C4-2 skeletal tumor tissues was analyzed by immunohistochemical staining using rabbit polyclonal antibodies against human survivin (Novus Biologicals; 1:300 dilution) and mouse monoclonal antibody against Ki67 (Dako; 1:100 dilution).

Techniques: Phospho-proteomics, Western Blot, Binding Assay, Luciferase, Activity Assay, Transfection, Expressing

Journal: Oncotarget

Article Title: Inhibition of skeletal growth of human prostate cancer by the combination of docetaxel and BKM1644: an aminobisphosphonate derivative

doi: 10.18632/oncotarget.8481

Figure Lengend Snippet: ( A ) Left panel: BKM1644 alone or combined with docetaxel significantly reduces serum PSA levels in athymic nude mice bearing C4-2 tumors; Right panel: statistical analysis on the in vivo effect of BKM1644. ( B ) BKM1644 treatment improves the bone architecture in C4-2 tumor-bearing mice. Red arrow: osteoblastic lesion; yellow arrow: osteolytic lesion. ( C ) BKM1644 inhibits the in vivo expression of survivin and Ki67, and induces apoptosis in C4-2 skeletal tumor, as demonstrated by IHC and TUNEL assays. Two-tailed t -test was used to calculate p values on the weighted index (W.I). *, ** indicate p -values < 0.05.

Article Snippet: Expression of survivin and Ki67 in C4-2 skeletal tumor tissues was analyzed by immunohistochemical staining using rabbit polyclonal antibodies against human survivin (Novus Biologicals; 1:300 dilution) and mouse monoclonal antibody against Ki67 (Dako; 1:100 dilution).

Techniques: In Vivo, Expressing, TUNEL Assay, Two Tailed Test