Journal: Oncology reports

Article Title: Inhibition of CD147 gene expression via RNA interference reduces tumor cell invasion, tumorigenicity and increases chemosensitivity to cisplatin in laryngeal carcinoma Hep2 cells.

doi: 10.3892/or.2010.1088

Figure Lengend Snippet: Figure 1. CD147 specific shRNAs results in the reduction of CD147 mRNA and protein levels in Hep2 cells. (A) Relative mRNA levels were analysed by quantitative RT-PCR. ß-actin was used as normalization control. *P<0.01 compared with Hep2. (B) Western blot analysis of CD147 protein expressions. ß-actin was used as loading control.

Article Snippet: Goat anti-mouse CD147 polyclonal antibody (1:50 dilution, Santa Cruz Biotechnology) was used in standard indirect immunoperoxidase procedures.

Techniques: Quantitative RT-PCR, Control, Western Blot

Journal: Oncology reports

Article Title: Inhibition of CD147 gene expression via RNA interference reduces tumor cell invasion, tumorigenicity and increases chemosensitivity to cisplatin in laryngeal carcinoma Hep2 cells.

doi: 10.3892/or.2010.1088

Figure Lengend Snippet: Figure 2. Proliferation of Hep2 cells stably expressing shRNA. Proliferation of Hep2 cells stably transfected with blank control (Hep2), pSilencer- CD147shRNA-control, pSilencer-CD147 shRNA1 and pSilencer-CD147 shRNA2 were analyzed by MTT assay as described earlier. Spectrometrics absorbance at 490 nm was measured using a microplate reader. Each group contained three wells. *P<0.01 compared with Hep2.

Article Snippet: Goat anti-mouse CD147 polyclonal antibody (1:50 dilution, Santa Cruz Biotechnology) was used in standard indirect immunoperoxidase procedures.

Techniques: Stable Transfection, Expressing, shRNA, Transfection, Control, MTT Assay

Journal: Oncology reports

Article Title: Inhibition of CD147 gene expression via RNA interference reduces tumor cell invasion, tumorigenicity and increases chemosensitivity to cisplatin in laryngeal carcinoma Hep2 cells.

doi: 10.3892/or.2010.1088

Figure Lengend Snippet: Figure 4. Effects of CD147 specific shRNAs on Matrigel invasion of Hep2 cells. (A) (a) Hep2; (b) Hep2/CD147 shRNA-control; (c) Hep2/CD147 shRNA1; (d) Hep2/CD147 shRNA2 (x400). (B) Number of cells migrated was evaluated in three fields for each experimental group and averaged. The average invaded cell number of the groups Hep2, Hep2/CD147shRNA- control, Hep2/CD147shRNA1 and Hep2/CD147shRNA2 was (63.37±2.53), (62.59±2.27), (24.21±1.36) and (16.74±1.58) respectively. Compared with Hep2, the cell invasiveness of Hep2/CD147shRNA1 and Hep2/CD147 shRNA2 was reduced by 38.20 and 26.41%, respectively *P<0.01.

Article Snippet: Goat anti-mouse CD147 polyclonal antibody (1:50 dilution, Santa Cruz Biotechnology) was used in standard indirect immunoperoxidase procedures.

Techniques: shRNA, Control

Journal: Oncology reports

Article Title: Inhibition of CD147 gene expression via RNA interference reduces tumor cell invasion, tumorigenicity and increases chemosensitivity to cisplatin in laryngeal carcinoma Hep2 cells.

doi: 10.3892/or.2010.1088

Figure Lengend Snippet: Figure 5. Effects of CD147 specific shRNAs on cisplatin sensitivity of Hep2 cells. Hep2, Hep2/CD147shRNA-control, Hep2/CD147shRNA1 and Hep2/CD147shRNA2 were treated with various concentrations of cisplatin. Cell viability was determined by MTT assay. Cell survival index was calculated as [A490(cisplatin+)/A490(cispatin-)].

Article Snippet: Goat anti-mouse CD147 polyclonal antibody (1:50 dilution, Santa Cruz Biotechnology) was used in standard indirect immunoperoxidase procedures.

Techniques: Control, MTT Assay

Journal: Oncology reports

Article Title: Inhibition of CD147 gene expression via RNA interference reduces tumor cell invasion, tumorigenicity and increases chemosensitivity to cisplatin in laryngeal carcinoma Hep2 cells.

doi: 10.3892/or.2010.1088

Figure Lengend Snippet: Figure 6. Effects of CD147 specific shRNAs on tumor formation in vivo and the expression of CD147 in tumor tissues. (A) Hep2, Hep2/CD147shRNA- control, Hep2/CD147shRNA1, Hep2/CD147shRNA2 cells were inoculated subcutaneously into the nude mice. Tumor growth was monitored and tumor volumes were calculated. *P<0.01, compared with Hep2 or Hep2/CD147 shRNA-control. (B) CD147 protein expression was determined by immuno- histochemistry staining in solid tumors derived from tumor tissues. (a) Hep2; (b) Hep2/CD147shRNA-control; (c) Hep2/CD147shRNA1; (d) Hep2/ CD147shRNA2 cells (x400). Differences in CD147 protein expression are shown. The data were obtained from three independent experiments. (C) Histological analysis was performed in implanted tumors with H&E staining. (a) Hep2; (b) Hep2/CD147shRNA-control; (c) Hep2/CD147 shRNA1; (d) Hep2/CD147shRNA2 cells (x400).

Article Snippet: Goat anti-mouse CD147 polyclonal antibody (1:50 dilution, Santa Cruz Biotechnology) was used in standard indirect immunoperoxidase procedures.

Techniques: In Vivo, Expressing, Control, shRNA, Immunohistochemistry, Staining, Derivative Assay

Journal: eLife

Article Title: Evolutionarily conserved sperm factors, DCST1 and DCST2, are required for gamete fusion

doi: 10.7554/eLife.66313

Figure Lengend Snippet: ( A ) Accumulation of many acrosome-reacted (AR) IZUMO1 positive spermatozoa (red) in the perivitelline space of eggs recovered from the oviducts of females 8 hr after coitus with a Dcst1/2 -/- male. Scale bar 10 µm. ( B ) Gamete fusion assay of Dcst1/2 -disrupted spermatozoa. Gamete fusion was visualized with Hoechst 33342 dye transfer. The numbers of fused spermatozoa per oocyte from four independent experiments are shown. The total numbers of oocytes examined in Dcst1/2 +/- and Dcst1/2 -/- were 77 and 80, respectively (left graph). The broken lines indicate the average. The arrows and the circles in the representative photo show fused spermatozoa and meiosis metaphase II chromosomes, respectively (right picture). Scale bar 10 µm. ( C ) Localization of oocyte CD9 and JUNO upon Dcst1/2 -/- sperm attachment. Sperm–oocyte interaction was visualized by staining with 0.25 µg ml −1 TH6–Alexa647 (JUNO: red), 0.5 µg ml −1 MZ3–FITC (CD9: green) and 0.5 µg ml −1 Mab125–Alexa546 (IZUMO1: blue). The eggs were fixed 30 min after insemination of Dcst1/2 -/- spermatozoa. The arrowheads indicate the sites where JUNO and cluster of differentiation 9 (CD9) were concentrated in response to the AR spermatozoa (IZUMO1 positive: blue) bound to the oolemma. Scale bar 10 µm. ( D ) In vitro fertilization assay using Dcst1/2 +/- , Dcst1/2 -/- , Dcst1 +/- , Dcst1 -/- , Dcst2 +/- , Dcst2 -/- , Dcst1/2 -/- Dcst1 -TG, Dcst1/2 -/- Dcst2 -TG, and Dcst1/2 -/- Dcst1/2 -TG spermatozoa (n = 5, 5, 6, 7, 8, 8, 6, 6, and 6, respectively). The fertilization rate was evaluated at the two-cell stage embryo. The numbers in parentheses indicate the numbers of oocytes used. The error bars represent s.e.m. ***p<0.001 (paired two-tailed Student’s t-test, compared with each heterozygous mutants). ( E ) Fecundity of DCST1/2-related deficient male and female mice. The numbers in parentheses indicate the numbers of mating pairs. Regarding Dcst1/2 -/- , Dcst1 -/- , Dcst2 -/- , Dcst1/2 -/- Dcst1 -TG, and Dcst1/2 -/- Dcst2 -TG males, the numbers of vaginal plug formations were counted. The error bars represent s.e.m. ( F ) Western blot analysis of various types of mouse lines with 1 µg ml −1 Mab18 (IZUMO1) and 1:500 SPACA6 anti-serum. Each lane was equally applied with 20 µg sperm lysates. BASIGIN was used as a loading control. Figure 3—source data 1. Gamete fusion assay, IVF and litter size in DCST1/2-related mutants.

Article Snippet: Antibody , Anti-mouse EMMPRIN (BASIGIN) (goat polyclonal) , Santa Cruz Biotechnology , G-19, RRID AB_2066959 , (WB: 0.2 μg ml −1 ).

Techniques: Single Vesicle Fusion Assay, Staining, In Vitro, Two Tailed Test, Western Blot, Control

Journal: eLife

Article Title: Evolutionarily conserved sperm factors, DCST1 and DCST2, are required for gamete fusion

doi: 10.7554/eLife.66313

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-mouse EMMPRIN (BASIGIN) (goat polyclonal) , Santa Cruz Biotechnology , G-19, RRID AB_2066959 , (WB: 0.2 μg ml −1 ).

Techniques: Recombinant, Plasmid Preparation, Sequencing

Journal:

Article Title: Non-Mitogenic Survival-Enhancing Autocrine Factors Including Cyclophilin A Contribute to Density-Dependent mESC Growth

doi: 10.1016/j.scr.2010.10.001

Figure Lengend Snippet: a. Image of a portion of a silver-stained gel resulting from SDS-PAGE of medium conditioned by ABJ1 cells and control (serum-free) medium. The arrows indicate the location of two additional bands observed in the ABJ1 CM lane. b. Fold growth on day 2 of mESCs cultured with different concentrations of cyclophilin A (added on day 2), showing that growth increases in a concentration-dependent manner (p=0.03) c. Fold growth on day 2 of mESCs after addition of no antibody, goat IgG, or a polyclonal antibody to CD147 (2 μg/ml), showing significantly decreased growth upon addition of the CD147 Ab ( p(CD147, normal IgG) = 4e-6, p(CD147, PBS) = 2e-8).

Article Snippet: Goat anti-mouse CD147 polyclonal antibody and goat IgG control were purchased from Santa Cruz Biotechnology.

Techniques: Staining, SDS Page, Cell Culture, Concentration Assay