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Effects of forced expression of ALDH1A2 on invasive potential of ovarian cancer cells. ( A , B ) Invasion assay; RMG-I cells transfected with either the empty vector or ALDH1A2 expression vector for 24 h were loaded into the upper chamber and then fetal bovine serum (FBS, 5%) was added to the lower chamber. After 24 h, the cells that invaded the lower chamber were stained with <t>DiffQuik</t> ( A ) and quantified ( B ) under a microscope. Data are expressed as means ± SD. Statistical significance was assessed using an unpaired t -test. *** p < 0.005. ( C ) Sprouting assay; RMG1 cells ectopically expressing ALDH1A2 were embedded with Matrigel and incubated for 24 h. Phase contrast images were observed (left panel). Calcein-AM-stained cells were visualized using a fluorescence microscope (right panel). Original magnification: 4×. Scale bar, 500 μm. ( D ) Sprouting assay using RMG1 cells incubated in the presence or absence of ATRA (1 μM, 5 μM). Phase contrast images were observed (left panel). Calcein-AM-stained cells were visualized using a fluorescence microscope (right panel). Original magnification: 4×. Scale bar, 500 μm.
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Effects of forced expression of ALDH1A2 on invasive potential of ovarian cancer cells. ( A , B ) Invasion assay; RMG-I cells transfected with either the empty vector or ALDH1A2 expression vector for 24 h were loaded into the upper chamber and then fetal bovine serum (FBS, 5%) was added to the lower chamber. After 24 h, the cells that invaded the lower chamber were stained with DiffQuik ( A ) and quantified ( B ) under a microscope. Data are expressed as means ± SD. Statistical significance was assessed using an unpaired t -test. *** p < 0.005. ( C ) Sprouting assay; RMG1 cells ectopically expressing ALDH1A2 were embedded with Matrigel and incubated for 24 h. Phase contrast images were observed (left panel). Calcein-AM-stained cells were visualized using a fluorescence microscope (right panel). Original magnification: 4×. Scale bar, 500 μm. ( D ) Sprouting assay using RMG1 cells incubated in the presence or absence of ATRA (1 μM, 5 μM). Phase contrast images were observed (left panel). Calcein-AM-stained cells were visualized using a fluorescence microscope (right panel). Original magnification: 4×. Scale bar, 500 μm.

Journal: Cancers

Article Title: ALDH1A2 Is a Candidate Tumor Suppressor Gene in Ovarian Cancer

doi: 10.3390/cancers11101553

Figure Lengend Snippet: Effects of forced expression of ALDH1A2 on invasive potential of ovarian cancer cells. ( A , B ) Invasion assay; RMG-I cells transfected with either the empty vector or ALDH1A2 expression vector for 24 h were loaded into the upper chamber and then fetal bovine serum (FBS, 5%) was added to the lower chamber. After 24 h, the cells that invaded the lower chamber were stained with DiffQuik ( A ) and quantified ( B ) under a microscope. Data are expressed as means ± SD. Statistical significance was assessed using an unpaired t -test. *** p < 0.005. ( C ) Sprouting assay; RMG1 cells ectopically expressing ALDH1A2 were embedded with Matrigel and incubated for 24 h. Phase contrast images were observed (left panel). Calcein-AM-stained cells were visualized using a fluorescence microscope (right panel). Original magnification: 4×. Scale bar, 500 μm. ( D ) Sprouting assay using RMG1 cells incubated in the presence or absence of ATRA (1 μM, 5 μM). Phase contrast images were observed (left panel). Calcein-AM-stained cells were visualized using a fluorescence microscope (right panel). Original magnification: 4×. Scale bar, 500 μm.

Article Snippet: After 24 h, the membranes were fixed with 100% methanol and stained with a DiffQuik stain kit (Triangle Biomedical Sciences, Inc., Durham, NC, USA).

Techniques: Expressing, Invasion Assay, Transfection, Plasmid Preparation, Staining, Microscopy, Incubation, Fluorescence