Journal: The Journal of Clinical Investigation

Article Title: HIF-2 α expression and metabolic signaling require ACSS2 in clear cell renal cell carcinoma

doi: 10.1172/JCI164249

Figure Lengend Snippet: ( A ) Left: Representative images of crystal violet staining of HEK293FT cells 72 hours after transfection with an empty vector control (EV), WT-ACSS2, or ΔT376K catalytically inactive mutant. Right: Bar graph showing quantification of 3 independent experiments in HEK293FT cells. Statistical significance was determined using a 1-way ANOVA and Tukey’s multiple-comparison test (* P < 0.05; ** P < 0.01). Data are represented as mean ± SD. ( B ) Western blot images showing expression of HIF-2α ( n = 3), ACSS2 ( n = 3), and actin ( n = 3) in 786-O Cas9 sgControl and sgACSS2 cells transfected with EV, WT-ACSS2, or ΔT376K. ( C ) Left: Representative images of crystal violet staining of 786-O Cas9 cells stably transduced to express sgControl or sgACSS2 72 hours after transfection with an empty vector control (EV), WT-ACSS2, or ΔT376K catalytically inactive mutant. Right: Bar graph showing quantification of 3 independent experiments in 786-O Cas9 cells. Statistical significance was determined using a 2-way ANOVA and Tukey’s multiple-comparison test (* P < 0.05; ** P < 0.01). Data are represented as mean ± SD. ( D ) Representative images from day 4 of a tumor sphere formation assay where 786-O Cas9 sgControl and sgACSS2 cells were transfected with EV, WT-ACSS2, or ΔT376K.

Article Snippet: 786-O Cas9 cells were purchased from Genecopoeia.

Techniques: Staining, Transfection, Plasmid Preparation, Control, Mutagenesis, Comparison, Western Blot, Expressing, Stable Transfection, Tube Formation Assay

Journal: International Journal of Molecular Sciences

Article Title: Regulated Mesenchymal Stem Cells Mediated Colon Cancer Therapy Assessed by Reporter Gene Based Optical Imaging

doi: 10.3390/ijms19041002

Figure Lengend Snippet: Bystander effect of MSC-Tet-TK and MSC-TK cells. ( A ) Rluc activity in co-cultures (1:1) of naive MSCs and CT26/Rluc cells treated with the indicated concentrations of GCV for 48 h. ( B ) BLI images of the Rluc activity and quantitation data of CT26/Rluc in co-cultures (1:1) of MSC-TK or MSC-Tet-TK cells in the absence or presence of doxycycline (DOX(−) and DOX 2 μg/mL respectively). Three individual experiment values are expressed as the mean ± standard deviation (SD), * p < 0.05, ** p < 0.01, *** p < 0.001 (by Student’s t test). p/s, photons/second.

Article Snippet: CT26 cells were transduced with lentiviral particles expressing mCherry-Rluc (Renilla luciferase) under the control of the cytomegalovirus (CMV) promoter (Genecopoeia, Rockville, MD, USA).

Techniques: Activity Assay, Quantitation Assay, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Regulated Mesenchymal Stem Cells Mediated Colon Cancer Therapy Assessed by Reporter Gene Based Optical Imaging

doi: 10.3390/ijms19041002

Figure Lengend Snippet: In vivo therapeutic effect of MSC-Tet-TK and MSC-TK cells on inhibiting colon tumor growth. ( A ) Renilla luciferase (Rluc) imaging of colon cancer cells (CT26/Rluc) in mice treated with either MSC-TK or MSC-Tet-TK cells with or without concurrent GCV treatment. BLI images were taken on days 0, 6, and 13 in five individual mice; ( B ) Quantitative analysis of the data shown in ( A ). ( C ) Tumor weights assessed at study end. Bioluminescence activity is shown in photons/second (p/s). * p < 0.05 compared separately to MSC-Tet-TK (GCV−) and MSC-TK (GCV−).

Article Snippet: CT26 cells were transduced with lentiviral particles expressing mCherry-Rluc (Renilla luciferase) under the control of the cytomegalovirus (CMV) promoter (Genecopoeia, Rockville, MD, USA).

Techniques: In Vivo, Luciferase, Imaging, Activity Assay

Journal: JCI Insight

Article Title: LIN28B induces a differentiation program through CDX2 in colon cancer

doi: 10.1172/jci.insight.140382

Figure Lengend Snippet: ( A ) Left: subcutaneous xenograft experiments with LoVo cells ( n = 6 per cell type) showed a significant increase in tumor weight with CDX2 KD with sh-CDX2 no. 1 (472.38 ± 126.23 mg at euthanization) or sh-CDX2 no. 2 (353.38 ± 112.90 mg at euthanization) as compared with control cells (123.88 ± 48.86 mg at euthanization). Right: the images of tumors. Scale bars: 10 mm. ( B ) Ki-67 staining in the subcutaneous xenograft tumor of LoVo cells with control or CDX2 KD, representative images (upper) and the quantification (lower). Scale bars: 100 μm. n = 3. ( C ) Transwell chamber invasion assay of LoVo cells with control or CDX2 KD, representative images (upper) and the quantification (lower). Scale bars: 100 μm. n = 3. ( D ) Ki-67 staining in the liver metastatic tumor of LoVo cells with control or CDX2 KD, representative images (upper) and the quantification (lower). Scale bars: 100 μm. n = 3. ( E ) Epithelial-mesenchymal transition (EMT) marker expression (qPCR) in LoVo control/CDX2 KD cells ( n = 3). ( F ) Representative IHC of CDX2, E-cadherin, and CK20 in the liver metastatic tumor of LoVo cells with control or CDX2 KD. Scale bars: 100 μm. Data are presented as mean ± SEM. Unpaired, 2-tailed Student’s t test ( D ) and 1-way ANOVA followed by Dunnett’s multiple-comparison test as post hoc analysis ( A , B , C , and E ) were performed. * P < 0.05, ** P < 0.01.

Article Snippet: CDX2 knockdown in Caco-2 cells and LoVo cells was performed using Genecopoeia shRNA system with HSH000553-31-LVRU6MH (shCDX2 no. 1) and HSH000553-33 LVRU6MH (shCDX no. 2) and CSHCTR001-LVRU6MH (sh-control) constructs.

Techniques: Control, Staining, Invasion Assay, Marker, Expressing, Comparison