Journal: bioRxiv

Article Title: Vector analysis of steerable mechanical tension across nuclear lamina

doi: 10.1101/462275

Figure Lengend Snippet: (A) MCF-7 cells were treated with ADM, H 2 O 2 , and CHX separately. The samples were stained for both β-actin and α-tubulin. The cells were also stained with DAPI to localize the nuclei. The images were captured through confocal laser microscopy after immunofluorescence staining (n = 10). (B) Intracellular nanoparticle size distribution in MCF-7 cells separately treated with ADM, H 2 O 2 , and CHX (n = 10). (C) MCF-7 cell intracellular osmotic pressure was measured via osmometery after the cells were exposed to ADM, H 2 O 2 , and CHX. (**: 0.001 < p < 0.05, ns: p > 0.05, Tukey-b test, n = 10). (D) P-cofilin, cofilin, actin, p-stathmin, stathmin, and tubulin levels in MCF-7 cells that had been treated with ADM, H 2 O 2 , or CHX (n = 6). (E) In the H 2 O 2 group, the ΔpNOP could be divided into two parts by comparing the total intensity of the protein nanoparticles, generated from MF and MT depolymerization, respectively (versus , line 1, row 2). Scale bar, 10 μm. All error bars represent SEM.

Article Snippet: Antibodies were purchased from commercial sources: rabbit anti-lamin B1 antibody (13435S, Cell Signaling Technology, Temecula, USA), rabbit anti-β-actin antibody (4970P, Cell Signaling Technology, Temecula, USA), mouse anti-tubulin-α antibody (T5168, Boster, Wuhan, China), rabbit anti-vimentin antibody (5741S, Cell Signaling Technology, Temecula, USA), rabbit anti-GFP antibody (2956S, Cell Signaling Technology, Temecula, USA), mouse anti-phospho-cofilin (Ser24) (bs-10252R, Bioss, Woburn, USA), mouse anti-cofilin antibody (PB0035, Boster, Wuhan, China), rabbit anti-phospho-stathmin 1 (Ser38) (bs-3432R, Bioss, Woburn, USA), and mouse anti-stathmin 1 antibody (BA1417, Boster, Wuhan, China).

Techniques: Staining, Microscopy, Immunofluorescence, Generated

Journal: Cancer Science

Article Title: Stathmin1 regulates p27 expression, proliferation and drug resistance, resulting in poor clinical prognosis in cholangiocarcinoma

doi: 10.1111/cas.12417

Figure Lengend Snippet: Clinicopathological characteristics of extrahepatic cholangiocarcinoma patients according to stathmin expression

Article Snippet: The sections were then heated in boiled water and Immunosaver (Nishin EM, Tokyo, Japan) at 98°C for 45 min. Non-specific binding sites were blocked by incubating with Protein Block Serum-Free (DAKO, Carpinteria, CA, USA) for 30 min. A mouse monoclonal anti-STMN1 (OP18) antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and a mouse monoclonal anti-p27 antibody (Santa Cruz Biotechnology) were applied at a dilution of 1:100 for 24 h at 4°C.

Techniques: Expressing, Labeling

Journal: Cancer Science

Article Title: Stathmin1 regulates p27 expression, proliferation and drug resistance, resulting in poor clinical prognosis in cholangiocarcinoma

doi: 10.1111/cas.12417

Figure Lengend Snippet: Univariate and multivariate analysis of prognostic factors using the Cox proportional hazards model

Article Snippet: The sections were then heated in boiled water and Immunosaver (Nishin EM, Tokyo, Japan) at 98°C for 45 min. Non-specific binding sites were blocked by incubating with Protein Block Serum-Free (DAKO, Carpinteria, CA, USA) for 30 min. A mouse monoclonal anti-STMN1 (OP18) antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and a mouse monoclonal anti-p27 antibody (Santa Cruz Biotechnology) were applied at a dilution of 1:100 for 24 h at 4°C.

Techniques: Adjuvant, Expressing

Journal: Cancer Science

Article Title: Stathmin1 regulates p27 expression, proliferation and drug resistance, resulting in poor clinical prognosis in cholangiocarcinoma

doi: 10.1111/cas.12417

Figure Lengend Snippet: (a) Stathmin1 (STMN1) bound to p27 in the cytoplasm of HuCCT1 cells. Primary STMN1 rabbit and p27 mouse antibodies bound to STMN1 and p27 complexes were combined with secondary proximity ligation assay probes. The interaction events are visible as red dots (nuclear staining with DAPI). (b) siRNA-mediated STMN1 suppression. STMN1 protein levels were measured using western blotting after transfection with STMN1 siRNA. p27 expression in STMN1 siRNA-transfected and untreated HuCCT-1 cells was assessed using western blotting of total and nuclear protein. β-Actin was used as a loading control for total protein and Histone H1 was used as a loading control for nuclear protein. (c) HuCCT-1 cells were transfected with STMN1 siRNA and WST-8 proliferative assays were performed and compared with untransfected cells. (d) Paclitaxel sensitivity was determined using a WST-8 assay. STMN1 siRNA transfection significantly increased paclitaxel sensitivity compared with untransfected cells. Each time point represents the mean ± SD of triplicate determinations in two independent experiments. * P < 0.05.

Article Snippet: The sections were then heated in boiled water and Immunosaver (Nishin EM, Tokyo, Japan) at 98°C for 45 min. Non-specific binding sites were blocked by incubating with Protein Block Serum-Free (DAKO, Carpinteria, CA, USA) for 30 min. A mouse monoclonal anti-STMN1 (OP18) antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and a mouse monoclonal anti-p27 antibody (Santa Cruz Biotechnology) were applied at a dilution of 1:100 for 24 h at 4°C.

Techniques: Proximity Ligation Assay, Staining, Western Blot, Transfection, Expressing, Control