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Image Search Results
Journal: Cancer cell
Article Title: Tinagl1 Suppresses Triple-Negative Breast Cancer Progression and Metastasis by Simultaneously Inhibiting Integrin/FAK and EGFR Signaling.
doi: 10.1016/j.ccell.2018.11.016
Figure Lengend Snippet: Figure 4. Identification of Tinagl1-Interacting Proteins (A–C) LM2 cells expressing the C-terminal HA-tagged Tinagl1 (Tinagl1-HA) were lysed and immunoprecipitated (IP) with immunoglobulin G (IgG) (control) or anti- HA antibody. The IP samples were subjected to silver staining and WB (A) before mass spectrometry analysis. Tinagl1-interacting partners were clustered with KEGG pathway analysis, and the three top pathways are shown in (B). The members of the top three pathways have overlaps. The EGFR and integrin b1 subunits are the core members of each pathways (C). (D and E) LM2 cells stably expressing Tinagl1-HA were lysed and IP with IgG or anti-HA antibodies. The IP samples were subjected to WB analysis with the indicated antibodies to detect the interaction with EGFR and the integrin b1 subunit (D), and with integrins av, or a5 subunits (E). See also Figure S4.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Tinagl1, in 1:1000 (WB), 2 mg (IP), Rabbit ProteinTech Cat#12077-1-AP; RRID: AB_2058942 Tinagl1, in 1:100 (IHC), Rabbit Sigma-Aldrich Cat#HPA048695; RRID: AB_2680497 b-actin, in 1:10,000 (WB), mouse Abcam Cat#ab6276; RRID: AB_2223210 EGFR, in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4267; RRID: AB_2246311 p-EGFR (Try1068), in 1:1000 (WB), 1:100 (IHC), Rabbit Cell Signaling Technology Cat#3777; RRID: AB_2096270 FAK, in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#3285; RRID: AB_10694068 p-FAK (Try397), in 1:1000 (WB), 1:100 (IHC), Rabbit Cell Signaling Technology Cat#8556; RRID: AB_10891442 p-FAK (Try925), in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#3284; RRID: AB_2253227 AKT, in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4691; RRID: AB_915783 p-ATK (S473), in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4060; RRID: AB_2315049 ERK1/2, in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4695; RRID: AB_390779 p-ERK1/2 (Thr202/Tyr204), in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4370; RRID: AB_2315112 Integrin b1 subunit, in 1:1000 (WB), 2 mg (IP), Rabbit Cell Signaling Technology Cat#34971
Techniques: Expressing, Immunoprecipitation, Control, Silver Staining, Mass Spectrometry, Stable Transfection
Journal: Cancer cell
Article Title: Tinagl1 Suppresses Triple-Negative Breast Cancer Progression and Metastasis by Simultaneously Inhibiting Integrin/FAK and EGFR Signaling.
doi: 10.1016/j.ccell.2018.11.016
Figure Lengend Snippet: Figure 5. Tinagl1 Inhibits integrin/FAK and EGFR Signaling Pathways (A) Gene set enrichment analysis of lung metastatic nodules formed by LM2 cells stably expressing the vector control or Tinagl1 were dissected and digested. n = 3 per group. (B) Heatmap representation of microarray data displaying the expression of EGFR or integrin/FAK regulated genes in the control versus Tinagl1-expressing LM2 cells. (C) Heatmap representation of microarray data displaying the expression of genes compensated by integrin/FAK (left) or EGFR (right) in control versus Tinagl1- expressing LM2 cells.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Tinagl1, in 1:1000 (WB), 2 mg (IP), Rabbit ProteinTech Cat#12077-1-AP; RRID: AB_2058942 Tinagl1, in 1:100 (IHC), Rabbit Sigma-Aldrich Cat#HPA048695; RRID: AB_2680497 b-actin, in 1:10,000 (WB), mouse Abcam Cat#ab6276; RRID: AB_2223210 EGFR, in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4267; RRID: AB_2246311 p-EGFR (Try1068), in 1:1000 (WB), 1:100 (IHC), Rabbit Cell Signaling Technology Cat#3777; RRID: AB_2096270 FAK, in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#3285; RRID: AB_10694068 p-FAK (Try397), in 1:1000 (WB), 1:100 (IHC), Rabbit Cell Signaling Technology Cat#8556; RRID: AB_10891442 p-FAK (Try925), in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#3284; RRID: AB_2253227 AKT, in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4691; RRID: AB_915783 p-ATK (S473), in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4060; RRID: AB_2315049 ERK1/2, in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4695; RRID: AB_390779 p-ERK1/2 (Thr202/Tyr204), in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4370; RRID: AB_2315112 Integrin b1 subunit, in 1:1000 (WB), 2 mg (IP), Rabbit Cell Signaling Technology Cat#34971
Techniques: Protein-Protein interactions, Stable Transfection, Expressing, Plasmid Preparation, Control, Microarray
Journal: Cancer cell
Article Title: Tinagl1 Suppresses Triple-Negative Breast Cancer Progression and Metastasis by Simultaneously Inhibiting Integrin/FAK and EGFR Signaling.
doi: 10.1016/j.ccell.2018.11.016
Figure Lengend Snippet: Figure 6. Tinagl1 Inhibits EGFR Dimerization and Blocks the Interaction between the Integrin b1 Subunit and FN (A) LM2 cells were transfected with plasmids to overexpress GFP-EGFR and EGFR-Myc. 48 hr after transfection, the cells were treated with or without 1 mg/mL of r-Tinagl1 for 1 hr, followed by 10 min of 1 ng/mL EGF treatment. The cells were then collected and immunoprecipitated with either IgG or anti-Myc antibody. IP samples were subjected to WB analysis (top), and the amount of EGFR-GFP that interacts with EGFR-Myc was quantified and normalized to the PBS treatment group (bottom). (B) LM2 cells stably expressing EGFR-Myc were pre-treated with PBS or 1 mg/mL of r-Tinagl1 for 1 hr and then treated with 1 ng/mL of EGF for another 10 min. Next, the cells were collected and the dimers were crosslinked with disuccinimidyl suberate (DSS) treatment, followed by WB analysis (top) and quantification of the ratio of dimerized EGFR in each treatment group (bottom). (C) HEK293T cells overexpressing the integrin b1 subunit were lysed. 20 mg of FN was added into the lysate, and the lysate was divided into eight groups. PBS or the indicated amount of proteins were added into each group followed by IP with IgG or anti-b1 antibody. The IP samples were then subjected to WB analysis. (D) HEK293T cells overexpressing both integrin b1 subunit and Tinagl1-HA were lysed and divided into eight groups. PBS or the indicated amount of proteins were added into the lysate, followed by IP and WB analysis.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Tinagl1, in 1:1000 (WB), 2 mg (IP), Rabbit ProteinTech Cat#12077-1-AP; RRID: AB_2058942 Tinagl1, in 1:100 (IHC), Rabbit Sigma-Aldrich Cat#HPA048695; RRID: AB_2680497 b-actin, in 1:10,000 (WB), mouse Abcam Cat#ab6276; RRID: AB_2223210 EGFR, in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4267; RRID: AB_2246311 p-EGFR (Try1068), in 1:1000 (WB), 1:100 (IHC), Rabbit Cell Signaling Technology Cat#3777; RRID: AB_2096270 FAK, in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#3285; RRID: AB_10694068 p-FAK (Try397), in 1:1000 (WB), 1:100 (IHC), Rabbit Cell Signaling Technology Cat#8556; RRID: AB_10891442 p-FAK (Try925), in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#3284; RRID: AB_2253227 AKT, in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4691; RRID: AB_915783 p-ATK (S473), in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4060; RRID: AB_2315049 ERK1/2, in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4695; RRID: AB_390779 p-ERK1/2 (Thr202/Tyr204), in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4370; RRID: AB_2315112 Integrin b1 subunit, in 1:1000 (WB), 2 mg (IP), Rabbit Cell Signaling Technology Cat#34971
Techniques: Transfection, Immunoprecipitation, Stable Transfection, Expressing
Journal: Cancer cell
Article Title: Tinagl1 Suppresses Triple-Negative Breast Cancer Progression and Metastasis by Simultaneously Inhibiting Integrin/FAK and EGFR Signaling.
doi: 10.1016/j.ccell.2018.11.016
Figure Lengend Snippet: Figure 7. Tinagl1 Inhibits TNBC Progression by Simultaneously Targeting the Integrin/FAK and EGFR Signaling Pathways (A) 104 LM2 cells was injected into the MFP of NSG mice. Mice were intravenously treated with the indicated reagents twice per week when tumors reached 2 mm in diameter. n = 6 mice per group. (B) WB analysis for the activation of EGFR and FAK in primary tumor of each group after 5 weeks of the treatments as in (A). (C) Quantification of tumor volumes of each treatment group of (A). n = 12 tumors per group. (D and E) Lungs were collected and spontaneous metastasis was examined by ex vivo BLI at the endpoint. n = 6 lungs per group. Representative lungs (D) and quantitative data (E) is shown. Data represent means ± SEM. n.s., not significant; p > 0.05, **p < 0.001, ***p < 0.0001. Significance determined by one-way ANOVA analysis with Dunnett’s test for multiple comparisons. See also Figure S7.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Tinagl1, in 1:1000 (WB), 2 mg (IP), Rabbit ProteinTech Cat#12077-1-AP; RRID: AB_2058942 Tinagl1, in 1:100 (IHC), Rabbit Sigma-Aldrich Cat#HPA048695; RRID: AB_2680497 b-actin, in 1:10,000 (WB), mouse Abcam Cat#ab6276; RRID: AB_2223210 EGFR, in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4267; RRID: AB_2246311 p-EGFR (Try1068), in 1:1000 (WB), 1:100 (IHC), Rabbit Cell Signaling Technology Cat#3777; RRID: AB_2096270 FAK, in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#3285; RRID: AB_10694068 p-FAK (Try397), in 1:1000 (WB), 1:100 (IHC), Rabbit Cell Signaling Technology Cat#8556; RRID: AB_10891442 p-FAK (Try925), in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#3284; RRID: AB_2253227 AKT, in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4691; RRID: AB_915783 p-ATK (S473), in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4060; RRID: AB_2315049 ERK1/2, in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4695; RRID: AB_390779 p-ERK1/2 (Thr202/Tyr204), in 1:1000 (WB), Rabbit Cell Signaling Technology Cat#4370; RRID: AB_2315112 Integrin b1 subunit, in 1:1000 (WB), 2 mg (IP), Rabbit Cell Signaling Technology Cat#34971
Techniques: Protein-Protein interactions, Injection, Activation Assay, Ex Vivo
Journal: Integrative zoology
Article Title: cba-miR-222-3p involved in photoperiod-induced apoptosis in testes of striped hamsters by targeting TRAF7.
doi: 10.1111/1749-4877.12918
Figure Lengend Snippet: Figure 3 Validation of cba-miR-222-3p targeting TRAF7 and TRAF7 expression in the testes of striped hamsters. (a) Sequences and peak maps of cba-miR-222-3p, TRAF7-WT, and TRAF7-MT. (b) Relative luciferase activity detected by Dual-Luciferase Reporter Assay. (c) Immunohistochemistry (IHC, tissue microarray [TMA]) of TRAF7 in testes. (d) Integrated density of TRAF7 detected by IHC (TMA; n = 4). (e) Protein expression levels of TRAF7 in the testes detected by western blot (n = 4). (f) Pearson correlation analysis of cba-miR-222-3p and TRAF7. LD, long daylength; MD, moderate daylength; SD, short daylength; ∗, P < 0.05; ∗∗, P < 0.01.
Article Snippet: After performing electrophoresis, the proteins were transferred to PVDF membranes, which were then incubated with
Techniques: Biomarker Discovery, Expressing, Luciferase, Activity Assay, Reporter Assay, Immunohistochemistry, Microarray, Western Blot
Journal: Integrative zoology
Article Title: cba-miR-222-3p involved in photoperiod-induced apoptosis in testes of striped hamsters by targeting TRAF7.
doi: 10.1111/1749-4877.12918
Figure Lengend Snippet: Figure 4 Expression of cba-miR-222-3p and TRAF7 and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) analysis after in vivo injection in the testes of striped hamsters. (a) Schematic diagram of in vivo injection experiment of miRNA mimics. (b) Fluorescence in situ hybridization (FISH, tissue microarray [TMA]) of cba-miR-222-3p in testes after in vivo injection. (c) Fluorescence intensity of cba-miR-222-3p detected by FISH (TMA; n = 4). (d) Immunohistochemistry (IHC, TMA) of TRAF7 in testes after in vivo injection. (e) Integrated density of TRAF7 detected by IHC (TMA; n = 4). (f) TUNEL (TMA) staining of the testes after in vivo injection. (g) The proportion of TUNEL (TMA) staining with apoptotic activity in the testes (n = 4). (h) Relative expression of MEKK3 (n = 6). (i) Relative expression of p38 (n = 6). (j) Relative expression of p53 (n = 6). AG, agomir injection; NC, agomir negative control injection; DAPI, 4′6′-diamidino-2-phenylindole. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001.
Article Snippet: After performing electrophoresis, the proteins were transferred to PVDF membranes, which were then incubated with
Techniques: Expressing, TUNEL Assay, In Vivo, Injection, Fluorescence, In Situ Hybridization, Microarray, Immunohistochemistry, Staining, Activity Assay, Negative Control
Journal: PLoS ONE
Article Title: Down Regulation of T Cell Receptor Expression in COPD Pulmonary CD8 Cells
doi: 10.1371/journal.pone.0071629
Figure Lengend Snippet: The 51 genes within the functional annotation cluster were input into the STRING database. A network of how each gene interacts with others was produced. A sub-group of genes which interact closely, primarily through binding to each other, was highlighted (inset image). The genes of interest were CD247; LCK: Lymphocyte-specific protein tyrosine kinase; LAT: Linker of activated T cells; SYK: spleen tyrosine kinase; VAV2: VAV2 guanine nucleotide exchange factor; VAV3: VAV3 guanine nucleotide exchange factor.
Article Snippet: TaqMan Reverse transcription PCR was performed in duplicate for each sample using 1 μl of cDNA in a 25 μl reaction in step 2 of the Verso 2-step QRT-PCR mix kit (Thermo-scientific) containing 0.5 μl of premade ABI TaqMac gene expression assays for CD247 (Hs00609525_m1, Applied Biosystems, Warrington, UK), Linker of activated T cells (LAT, Hs01065378_g1, Applied Biosystems), LCK (Hs00178427_m1, Applied Biosystems), VAV2 guanine nucleotide exchange factor (VAV2,
Techniques: Functional Assay, Produced, Binding Assay
Journal: PLoS ONE
Article Title: Down Regulation of T Cell Receptor Expression in COPD Pulmonary CD8 Cells
doi: 10.1371/journal.pone.0071629
Figure Lengend Snippet: Gene expression levels of CD247, Leukocyte-specific protein tyrosine kinase (LCK); Linker of activated T cells (LAT); VAV2 guanine nucleotide exchange factor (VAV2) and VAV3 guanine nucleotide exchange factor (VAV3) were measured by both human U133 plus 2 affymetrix gene assay and by quantitative reverse transcription polymerase chain reaction (PCR). Fold change from blood to lung is charted. Statistically significant differences between blood and pulmonary levels measured by PCR are indicated by *p<0.05, **p<0.001. Statistically significant differences between blood and pulmonary levels measured by microarray are indicated by † Q<0.01.
Article Snippet: TaqMan Reverse transcription PCR was performed in duplicate for each sample using 1 μl of cDNA in a 25 μl reaction in step 2 of the Verso 2-step QRT-PCR mix kit (Thermo-scientific) containing 0.5 μl of premade ABI TaqMac gene expression assays for CD247 (Hs00609525_m1, Applied Biosystems, Warrington, UK), Linker of activated T cells (LAT, Hs01065378_g1, Applied Biosystems), LCK (Hs00178427_m1, Applied Biosystems), VAV2 guanine nucleotide exchange factor (VAV2,
Techniques: Gene Expression, Gene Assay, Reverse Transcription, Polymerase Chain Reaction, Microarray
Journal: PLoS ONE
Article Title: Down Regulation of T Cell Receptor Expression in COPD Pulmonary CD8 Cells
doi: 10.1371/journal.pone.0071629
Figure Lengend Snippet: Pulmonary CD8 cells were isolated from chronic obstructive pulmonary disease (COPD, grey n = 6) and smokers with normal lung function (S, black, n = 6). Transcript levels of CD247, Leukocyte-specific protein tyrosine kinase (LCK); Linker of activated T cells (LAT); VAV2 guanine nucleotide exchange factor (VAV2) and VAV3 guanine nucleotide exchange factor (VAV3) were measured by quantitative real time polymerase chain reaction. Statistically significant differences between COPD and S are represented by *p<0.05.
Article Snippet: TaqMan Reverse transcription PCR was performed in duplicate for each sample using 1 μl of cDNA in a 25 μl reaction in step 2 of the Verso 2-step QRT-PCR mix kit (Thermo-scientific) containing 0.5 μl of premade ABI TaqMac gene expression assays for CD247 (Hs00609525_m1, Applied Biosystems, Warrington, UK), Linker of activated T cells (LAT, Hs01065378_g1, Applied Biosystems), LCK (Hs00178427_m1, Applied Biosystems), VAV2 guanine nucleotide exchange factor (VAV2,
Techniques: Isolation, Real-time Polymerase Chain Reaction
Journal: International Journal of Oncology
Article Title: Carbon ion irradiation of the human prostate cancer cell line PC3: A whole genome microarray study
doi: 10.3892/ijo.2014.2287
Figure Lengend Snippet: The Applied Biosystems assays.
Article Snippet: ROCK1 , Rho-associated; coiled-coil containing protein kinase 1 ,
Techniques:
Journal: International Journal of Oncology
Article Title: Carbon ion irradiation of the human prostate cancer cell line PC3: A whole genome microarray study
doi: 10.3892/ijo.2014.2287
Figure Lengend Snippet: Significantly regulated motility genes: FC ≤-2 or FC ≥2 and FDR ≤0.05.
Article Snippet: ROCK1 , Rho-associated; coiled-coil containing protein kinase 1 ,
Techniques:
Journal: International Journal of Oncology
Article Title: Carbon ion irradiation of the human prostate cancer cell line PC3: A whole genome microarray study
doi: 10.3892/ijo.2014.2287
Figure Lengend Snippet: Microarray data of motility genes selected for RT-qPCR confirmation.
Article Snippet: ROCK1 , Rho-associated; coiled-coil containing protein kinase 1 ,
Techniques: Microarray
Journal: International Journal of Oncology
Article Title: Carbon ion irradiation of the human prostate cancer cell line PC3: A whole genome microarray study
doi: 10.3892/ijo.2014.2287
Figure Lengend Snippet: Relative gene expression changes of six selected motility genes 8 h after carbon ion or X-irradiation. Log 2 (ratio) of the expression of (A) MYH9 , s(B) ROCK1 , (C) NEXN , (D) FN1 , (E) MYH10 and (F) CCDC88A is presented. Significantly altered gene expression compared to CTRL samples ( * P≤0.05) based on one-tailed Mann-Whitney tests. RT-qPCR results confirm the downregulation observed by microarray analysis after radiation which was more pronounced after carbon ion radiation when compared to X-rays.
Article Snippet: ROCK1 , Rho-associated; coiled-coil containing protein kinase 1 ,
Techniques: Gene Expression, Irradiation, Expressing, One-tailed Test, MANN-WHITNEY, Quantitative RT-PCR, Microarray
Journal: International Journal of Oncology
Article Title: Carbon ion irradiation of the human prostate cancer cell line PC3: A whole genome microarray study
doi: 10.3892/ijo.2014.2287
Figure Lengend Snippet: Kaplan-Meier survival analysis of (A) CCDC88A , (B) FN1 , (C) MYH9 , (D) MYH10 , (E) NEXN and (F) ROCK1 gene expression performed on the data set of Taylor et al . Tumor samples were divided into three groups based on whether the gene expression value was high (♦, dark grey); intermediate (▴, light grey); or low (•, black). Differences in survival were found to be significant for CCDC88A , FN1 , NEXN and ROCK1 when log-rank P≤0.05.
Article Snippet: ROCK1 , Rho-associated; coiled-coil containing protein kinase 1 ,
Techniques: Gene Expression
Journal: International Journal of Oncology
Article Title: Carbon ion irradiation of the human prostate cancer cell line PC3: A whole genome microarray study
doi: 10.3892/ijo.2014.2287
Figure Lengend Snippet: Kaplan-Meier survival analysis of (A) CCDC88A , (B) FN1 , (C) MYH9 , (D) MYH10 , (E) NEXN and (F) ROCK1 gene expression performed on the data set of Gulzar et al . Tumor samples were divided into three groups based on whether the gene expression value was high (♦, dark grey); intermediate (▴, light grey); or low (•, black). Differences in survival were found to be significant for FN1 (log-rank P=0.0257).
Article Snippet: ROCK1 , Rho-associated; coiled-coil containing protein kinase 1 ,
Techniques: Gene Expression