Journal: Journal of Zhejiang University SCIENCE B

Article Title: ST13, a proliferation regulator, inhibits growth and migration of colorectal cancer cell lines

doi: 10.1631/jzus.b1200037

Figure Lengend Snippet: Fig. 1 ST13 expression in colorectal cancer cell lines (a) qRT-PCR shows relative transcript levels of ST13 in six colorectal cancer cell lines, including RKO, HT29, SW480, SW620, LOVO, and LS174T, and HEK293. The relative quantity was normalized to HEK293 cells, and β-actin was used as an internal control. (b) qRT-PCR shows relative transcript levels of ST13 in ST13-lentivirus-infected Lenti- ST13, control lentivirus-infected Lenti-Mock, and SW620. The relative quantity was normalized to Lenti-Mock cells, and β-actin was used as an internal control. (c) qRT-PCR shows relative transcript levels of ST13 in SW620, control- shRNA-lentivirus-infected shRNA-Mock, and ST13-shRNA- lentivirus-infected shRNA-ST13. The relative quantity was normalized to shRNA-Mock cells, and β-actin was used as an internal control. (d) Western blot results show that ST13 was expressed in SW620 cells with different groups, including Lenti-ST13, Lenti-Mock, SW620, shRNA-Mock, and shRNA-ST13. β-actin was served as loading control. Error bars indicate SEM (n=3 experiments)

Article Snippet: 2.3 Establishment of stable ST13 knockdown SW620 cell clones Small hairpin RNA (shRNA) lentiviral particles used for ST13 knockdown (sc-40684-v) and Mock knockdown (sc-108080) were purchased from Santa Cruz, CA, USA.

Techniques: Expressing, Quantitative RT-PCR, Control, Infection, shRNA, Western Blot

Journal: Journal of Zhejiang University SCIENCE B

Article Title: ST13, a proliferation regulator, inhibits growth and migration of colorectal cancer cell lines

doi: 10.1631/jzus.b1200037

Figure Lengend Snippet: Fig. 4 Effect of ST13 expression on SW620 cells mi- gration in vitro Lenti-ST13, Lenti-Mock, SW620, shRNA-Mock, and shRNA-ST13 cells were cultured in the top well of a transwell system and migration of SW620 cells was measured by counting the number of SW620 cells migrat- ing to the filter surface. (a) Quantitative measurement of invaded cells. Data are representatives of each group and expressed as mean±SEM of cells per six high power fields from three separated experiments. ** P<0.01, Lenti-ST13 vs. Lenti-Mock and SW620 (one way ANOVA and Dun- nett’s test). (b) Images of Lenti-ST13, Lenti-Mock, SW620, shRNA-Mock, and shRNA-ST13 cells on the filter surface

Article Snippet: 2.3 Establishment of stable ST13 knockdown SW620 cell clones Small hairpin RNA (shRNA) lentiviral particles used for ST13 knockdown (sc-40684-v) and Mock knockdown (sc-108080) were purchased from Santa Cruz, CA, USA.

Techniques: Expressing, In Vitro, shRNA, Cell Culture, Migration

Journal: Journal of Zhejiang University SCIENCE B

Article Title: ST13, a proliferation regulator, inhibits growth and migration of colorectal cancer cell lines

doi: 10.1631/jzus.b1200037

Figure Lengend Snippet: Fig. 5 Effect of ST13 expression on the development and growth of inoculated SW620 cells BALB/c nude mice (n=4 per group) were inoculated with 5×106 shRNA-ST13 or shRNA-Mock cells and the devel- opment of solid SW620 tumors was monitored every 3‒4 d. The mice were sacrificed 32 d post-inoculation and the tumors were taken at the same time. (a) The dynamics of shRNA-ST13 and shRNA-Mock tumor growth. Data are expressed as mean±SEM for each group. (b) The images of individual tumors

Article Snippet: 2.3 Establishment of stable ST13 knockdown SW620 cell clones Small hairpin RNA (shRNA) lentiviral particles used for ST13 knockdown (sc-40684-v) and Mock knockdown (sc-108080) were purchased from Santa Cruz, CA, USA.

Techniques: Expressing, shRNA

Journal:

Article Title: Overgrowth of a mouse model of Simpson- Golabi-Behmel syndrome is partly mediated by Indian Hedgehog

doi: 10.1038/embor.2009.98

Figure Lengend Snippet: Characterization and consequences of GPC3–lhh interaction. (A–C) Ihh binds to GPC3. (A) Binding of 125I-Ihh to human embryonic kidney (HEK)293T cells transfected with GPC3 expressing vector or vector controls (EF). A representative experiment of three is shown: points, average of triplicates; bars, ±s.d. (B) The specific binding of 125I-Ihh to HEK293T cells transfected with GPC3 or EF was determined in the presence of 100 × unlabelled Ihh. Bars: average of triplicates+s.d.; asterisk: statistically significant difference. (C) SRP analysis of the GPC3–Ihh interaction. GPC3ΔGPI was immobilized in the flow cell into streptavidin-coated SA sensor chips, and various concentrations of Ihh (bottom to top: 15, 30, 60, 120 and 240 nM) were injected on the surface of flow cells. The nonspecific binding was subtracted from the sensogram. Inset: ka, kd and Kd values were determined using a 1:1 Languimuir binding model. Each value is expressed as the mean±s.e. of five different concentrations. (D) GPC3 inhibits Hh signalling independently of CDO and HIP. Top: NIH 3T3 cells were transfected with a GPC3 or EF along with a luciferase reporter vector driven by an Hh responsive promoter (8Xgli) and β-galactosidase. As indicated, siRNA for silencing CDO siRNA, HIP siRNA or non-targeting control siRNA was also transfected. Cells were stimulated for 48 h with Shh- or control-conditioned medium and luciferase and β-galactosidase assays carried out. For each siRNA condition, the fold stimulation induced by Hh in EF-transfected cells was considered to be 100%. Bars represent the luciferase activity as percentage of the control (average+s.d. of triplicates). Bottom: Western blot for CDO and HIP in NIH 3T3 cells treated as indicated, using actin as loading control. (E,F) Increased Ihh levels in GPC3-null mice. (E) Western blot analysis of Ihh levels in whole-embryo lysates using actin as loading control was carried out. Bands were then scanned and quantified by densitometry. Average density+s.d. of Ihh/actin ratio is shown. In all three independent litters were analysed. (F) Relative levels of Ihh transcripts determined by real-time PCR using β-actin transcript levels as reference. Two independent litters were analysed. Bars represent mean+s.d. for the indicated genotypes. CDO, CAM-related/downregulated by oncogenes; EF, elongation factor; GPC3, Glypican 3; Hh, hedgehog; HIP, Hedgehog-interacting protein; Ihh, Indian hedgehog; mRNA, messenger RNA; PCR, polymerase chain reaction; RU, relative units; Shh, sonic hedgehog; siRNA, small interference RNA; SRP, small plasmon resonance; WT, wild type.

Article Snippet: To silence CDO or HIP, 100 nM of CDO siRNA (sc-60346, Santa Cruz), HIP siRNA (sc-40164, Santa Cruz) or non-targeting control siRNA A (sc-37007, Santa Cruz) was transfected using DharmaFECT-1 transfection reagent (Dharmacon, Chicago, IL, USA). β-Galactosidase activity was used to normalize the transfection efficiencies.

Techniques: Binding Assay, Transfection, Expressing, Plasmid Preparation, Injection, Luciferase, Control, Activity Assay, Western Blot, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction