Journal: Molecular medicine reports

Article Title: A systematic study of Girdin on cell proliferation, migration and angiogenesis in different breast cancer subtypes.

doi: 10.3892/mmr.2017.6971

Figure Lengend Snippet: Figure 3. Effect of Girdin knockdown on viability in breast cancer cells. (A) MCF‑7, (B) T47D and (C) MDA‑MB‑231 cells were transfected with Girdin siRNA or a negative control siRNA, and 48 h post‑transfection, viability was examined by a colorimetric MTT assay. Cells were also photographed under a phase‑contrast microscope at x400 magnification (left panels). Data are expressed as the mean ± standard deviation (n=3). **P<0.01 vs. control siRNA‑transfected cells. siRNA, small interfering RNA; MTT, 3‑(4,5‑Dimethylthiazol‑2‑yl)‑2,5‑diphenyltertrazolium bromide.

Article Snippet: Girdin siRNA (sc-94984) and non-targeting negative control siRNA (SIC002) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA) and GenePharma Co., Ltd. (Shanghai, China) respectively.

Techniques: Knockdown, Transfection, Negative Control, MTT Assay, Microscopy, Standard Deviation, Control, Small Interfering RNA

Journal: Molecular medicine reports

Article Title: A systematic study of Girdin on cell proliferation, migration and angiogenesis in different breast cancer subtypes.

doi: 10.3892/mmr.2017.6971

Figure Lengend Snippet: Figure 2. Efficiency of Girdin knockdown by siRNA. (A) MCF‑7, (B) T47D and (C) MDA‑MB‑231 cells were transfected with either a Girdin‑targeting siRNA or a non‑targeting negative control siRNA, and mRNA expression was examined using the reverse transcription‑semi‑quantitative polymerase chain reaction. 18S was used an internal reference for normalization. Data are expressed as the mean relative to control ± standard deviation (n=3). **P<0.01 vs. control siRNA‑transfected cells. siRNA, small interfering RNA; 18S, 18S ribosomal RNA.

Article Snippet: Girdin siRNA (sc-94984) and non-targeting negative control siRNA (SIC002) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA) and GenePharma Co., Ltd. (Shanghai, China) respectively.

Techniques: Knockdown, Transfection, Negative Control, Expressing, Polymerase Chain Reaction, Control, Standard Deviation, Small Interfering RNA

Journal: Molecular medicine reports

Article Title: A systematic study of Girdin on cell proliferation, migration and angiogenesis in different breast cancer subtypes.

doi: 10.3892/mmr.2017.6971

Figure Lengend Snippet: Figure 1. The structure of Human Girdin. A schematic for the proposed primary structure of the human Girdin protein. A dimerization domain is located at the N‑terminus, a large coiled‑coil domain at the central portion of the protein, and an actin‑binding domain at the C‑terminus. Akt phosphory lates Girdin at Serine 1416 (approximate location depicted by the arrow). Adapted from Jiang et al (4). Akt, RAC‑α serine/threonine‑protein kinase.

Article Snippet: Girdin siRNA (sc-94984) and non-targeting negative control siRNA (SIC002) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA) and GenePharma Co., Ltd. (Shanghai, China) respectively.

Techniques:

Journal: Molecular medicine reports

Article Title: A systematic study of Girdin on cell proliferation, migration and angiogenesis in different breast cancer subtypes.

doi: 10.3892/mmr.2017.6971

Figure Lengend Snippet: Figure 5. Effect of Girdin knockdown on VEGF expression in breast cancer cells. (A) MCF‑7, (B) T47D and (C) MDA‑MB‑231 cells were transfected with Girdin siRNA or a negative control siRNA, and then VEGF mRNA expres sion levels were examined using the reverse transcription‑semi‑quantitative polymerase chain reaction. 18S was used an internal reference for normaliza tion. Data are expressed as the mean ± standard deviation (n=3). **P<0.01 vs. control siRNA‑transfected cells. VEGF, vascular endothelial growth factor; siRNA, small interfering RNA; 18S, 18S ribosomal RNA.

Article Snippet: Girdin siRNA (sc-94984) and non-targeting negative control siRNA (SIC002) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA) and GenePharma Co., Ltd. (Shanghai, China) respectively.

Techniques: Knockdown, Expressing, Transfection, Negative Control, Polymerase Chain Reaction, Standard Deviation, Control, Small Interfering RNA

Journal: Molecular medicine reports

Article Title: A systematic study of Girdin on cell proliferation, migration and angiogenesis in different breast cancer subtypes.

doi: 10.3892/mmr.2017.6971

Figure Lengend Snippet: Figure 4. Effect of Girdin knockdown on migration in breast cancer cells. (A) MCF‑7, (B) T47D and (C) MDA‑MB‑231 cells were transfected with Girdin siRNA or a negative control siRNA, and then migration ability was examined using a chamber migration transwell assay. Four non‑overlapping fields per filter were selected and the migrated cells were counted. The average number of the cells from four fields was presented as the results of the migration assay. Representative images of the migrated cells at the bottom of the transwell filters are shown in the left panels (magnification, x100). Data are expressed as the mean ± standard deviation (n=3). **P<0.01 vs. control siRNA‑transfected cells. siRNA, small interfering RNA.

Article Snippet: Girdin siRNA (sc-94984) and non-targeting negative control siRNA (SIC002) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA) and GenePharma Co., Ltd. (Shanghai, China) respectively.

Techniques: Knockdown, Migration, Transfection, Negative Control, Transwell Assay, Standard Deviation, Control, Small Interfering RNA

Journal: Molecular medicine reports

Article Title: A systematic study of Girdin on cell proliferation, migration and angiogenesis in different breast cancer subtypes.

doi: 10.3892/mmr.2017.6971

Figure Lengend Snippet: Figure 7. Schematic model describing the potential molecular mechanism of Girdin action in breast cancer cells. Girdin knockdown suppressed cell viability, migration and angiogenesis in the three different breast cancer subtypes examined in the present study (breast epithelial cancer, breast ductal carcinoma and metastatic breast cancer), potentially by downregu lating the PI3K/Akt signaling pathway. PI3K, phosphatidyl inositol 3‑kinase; Akt, RAC‑α serine/threonine‑protein kinase.

Article Snippet: Girdin siRNA (sc-94984) and non-targeting negative control siRNA (SIC002) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA) and GenePharma Co., Ltd. (Shanghai, China) respectively.

Techniques: Knockdown, Migration

Journal: Molecular medicine reports

Article Title: A systematic study of Girdin on cell proliferation, migration and angiogenesis in different breast cancer subtypes.

doi: 10.3892/mmr.2017.6971

Figure Lengend Snippet: Figure 6. Effect of Girdin knockdown on protein expression of PI3K and Akt. MCF‑7, T47D and MDA‑MB‑231 cells were transfected with Girdin siRNA or a negative control siRNA, and then PI3K and Akt protein expres sion levels were examined by western bolt analysis, with β‑actin as the normalization reference. (A) Representative blot images. (B) Quantification for PI3K. (C) Quantification for Akt. Data are expressed as the mean relative to control ± standard deviation (n=3). **P<0.01 vs. control siRNA‑transfected cells. PI3K, phosphatidyl inositol 3‑kinase; Akt, RAC‑α serine/threo nine‑protein kinase; siRNA, small interfering RNA.

Article Snippet: Girdin siRNA (sc-94984) and non-targeting negative control siRNA (SIC002) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA) and GenePharma Co., Ltd. (Shanghai, China) respectively.

Techniques: Knockdown, Expressing, Transfection, Negative Control, Western Blot, Control, Standard Deviation, Small Interfering RNA

Journal: Frontiers in Oncology

Article Title: Inositol Polyphosphate 4-Phosphatase Type II Is a Tumor Suppressor in Multiple Myeloma

doi: 10.3389/fonc.2021.785297

Figure Lengend Snippet: Expression of INPP4B in MM patient samples. (A) The expression level of INPP4B in newly diagnosed MM patient samples. (B) Correlation of INPP4B expression with progression-free survival (PFS) and (C) overall survival (OS) in patients with newly diagnoses MM patients.

Article Snippet: After blocking, appropriately diluted primary INPP4B (#14543, Cell Signaling Technology) antibodies were added onto the slides and incubated in a humidified chamber at 4°C overnight, and then, appropriately diluted biotinylated secondary antibody was incubated at room temperature for 1 h. 3,3′-Diaminobenzidine (DAB) substrate solution (Dako, K5361) (freshly made just before use) was used to reveal the color of antibody staining.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: Inositol Polyphosphate 4-Phosphatase Type II Is a Tumor Suppressor in Multiple Myeloma

doi: 10.3389/fonc.2021.785297

Figure Lengend Snippet: Expression of INPP4B in EMP samples and cell lines. (A) Immunohistochemical images of EMP tissue (focal positive: a, b, c; negative: d, e, f). Scar = 50 μm. (B) Patients stratified according to their INPP4B expression level (positive and negative) and correlation of INPP4B expression with progression-free survival (PFS). (C) Overall survival (OS) rate of patients with these extramedullary involvements. (D) Correlation of INPP4B expression and overall survival rate of MM patients in GSE7982 cohort. (E) INPP4B protein expression in various multiple myeloma cell lines. (F) INPP4B mRNA expression in various multiple myeloma cell lines. Data are plotted as mean ± SD from three independent experiments (** p < 0.01, vs. the control).

Article Snippet: After blocking, appropriately diluted primary INPP4B (#14543, Cell Signaling Technology) antibodies were added onto the slides and incubated in a humidified chamber at 4°C overnight, and then, appropriately diluted biotinylated secondary antibody was incubated at room temperature for 1 h. 3,3′-Diaminobenzidine (DAB) substrate solution (Dako, K5361) (freshly made just before use) was used to reveal the color of antibody staining.

Techniques: Expressing, Immunohistochemical staining, Control

Journal: Frontiers in Oncology

Article Title: Inositol Polyphosphate 4-Phosphatase Type II Is a Tumor Suppressor in Multiple Myeloma

doi: 10.3389/fonc.2021.785297

Figure Lengend Snippet: INPP4B overexpression inhibits the proliferation of MM cells in vitro . (A) Western blot shows the overexpression of INPP4B in LP-1, RPMI8226, and MM.1S cells infected with lentivirus carrying INPP4B. (B) Western blot assay reveals the efficiency of INPP4B knockdown by lentiviral-carrying infection of shRNA. The CCK8 assay to detect the effect of INPP4B knockdown (KD) and overexpression (OE) on cells proliferation in (C) LP-1, (D) RPMI8226, and (E) MM.1S cells. NC, non-target control. (F) Cell cycle assay shows that overexpression of INPP4B arrested MM cells at G0/G1 phase. Data were plotted as mean ± SD from three independent experiments (* p < 0.05, ** p < 0.01, *** p < 0.001 vs. the control).

Article Snippet: After blocking, appropriately diluted primary INPP4B (#14543, Cell Signaling Technology) antibodies were added onto the slides and incubated in a humidified chamber at 4°C overnight, and then, appropriately diluted biotinylated secondary antibody was incubated at room temperature for 1 h. 3,3′-Diaminobenzidine (DAB) substrate solution (Dako, K5361) (freshly made just before use) was used to reveal the color of antibody staining.

Techniques: Over Expression, In Vitro, Western Blot, Infection, Knockdown, shRNA, CCK-8 Assay, Control, Cell Cycle Assay

Journal: Frontiers in Oncology

Article Title: Inositol Polyphosphate 4-Phosphatase Type II Is a Tumor Suppressor in Multiple Myeloma

doi: 10.3389/fonc.2021.785297

Figure Lengend Snippet: INPP4B inhibits MM cells growth via regulating the PI3K/Akt pathway. (A) Western blot analyzes the relative protein levels of Akt, p-Akt (thr308), p-Akt (ser473), rictor-mTOR complex (mTORC2) in LP-1, RPMI 8226, and MM.1S cells after lentiviral-carrying INPP4B overexpression for 48 h and (B) detections in LP-1, RPMI 8226, and MM.1S cells after lentiviral-carrying INPP4B knockdown for 48 h.

Article Snippet: After blocking, appropriately diluted primary INPP4B (#14543, Cell Signaling Technology) antibodies were added onto the slides and incubated in a humidified chamber at 4°C overnight, and then, appropriately diluted biotinylated secondary antibody was incubated at room temperature for 1 h. 3,3′-Diaminobenzidine (DAB) substrate solution (Dako, K5361) (freshly made just before use) was used to reveal the color of antibody staining.

Techniques: Western Blot, Over Expression, Knockdown

Journal: Frontiers in Oncology

Article Title: Inositol Polyphosphate 4-Phosphatase Type II Is a Tumor Suppressor in Multiple Myeloma

doi: 10.3389/fonc.2021.785297

Figure Lengend Snippet: INPP4B promotes chemosensitivity to proteasome inhibitor in MM cells. (A) Flow cytometry assay showed the apoptosis of MM.1S cells infected with lentiviral-carrying INPP4B expressing vector and treated with different dosage of bortezomib (BTZ) for 48 h, and (B) showed the statistical analysis for three independent experiments. (C) Flow cytometry assay showed the apoptosis of MM.1S cells infected with lentiviral-carrying shRNA targeting INPP4B and treated with different dosage of bortezomib (BTZ) for 48 h, and (B) showed the statistical analysis for three independent experiments.

Article Snippet: After blocking, appropriately diluted primary INPP4B (#14543, Cell Signaling Technology) antibodies were added onto the slides and incubated in a humidified chamber at 4°C overnight, and then, appropriately diluted biotinylated secondary antibody was incubated at room temperature for 1 h. 3,3′-Diaminobenzidine (DAB) substrate solution (Dako, K5361) (freshly made just before use) was used to reveal the color of antibody staining.

Techniques: Flow Cytometry, Infection, Expressing, Plasmid Preparation, shRNA