Cell colony formation assay was employed to evaluate cell proliferation. C666-1, CNE-2Z and HNE-2Z cells were plated into 6-well culture plates at a density of 1x103 cells per well and then treated with ZLMP1-C or ZWT affibody molecule for 14 days. Next, cells were washed with pre-cooled PBS, fixed with 4% paraformaldehyde and then stained with 0.1% Crystal Violet Staining Solution (Amresco, Solon, OH, United States). The number of colonies greater than 50 were counted under a microscope.
Crystal violet staining solution
Manufactured by Avantor
Sourced in United States
Crystal Violet Staining Solution is a laboratory reagent used for staining and visualization of cells or microorganisms. It is a component in various staining procedures, such as Gram staining, to enhance contrast and identification under a microscope.
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3 protocols using crystal violet staining solution
1
Cell Proliferation Assessment via Colony Assay
2
NPC Cell Proliferation Assay
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The cell proliferation of NPC cells was evaluated using a colony formation assay. Briefly, C666-1, CNE-2Z, and HNE-2 cells were seeded at (1 × 103 cells/well) in 6-well plates and then treated with ZLMP110-277, ZLMP277-110, ZLMP1-C277, or ZLMP2A-N110 (100 μg/mL) for 14 days. Subsequently, cells were washed with PBS, fixed with 4% paraformaldehyde, and stained with 0.1% Crystal Violet Staining Solution (Amresco, Solon, OH, USA). A light microscope was used to count the colonies formed (a group of more than 50).
3
Transwell Invasion Assay Protocol
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Following harvesting, 200 µl cell suspension containing 2×104 cells (resuspended in serum-free culture medium) were seeded in upper Transwell chambers (Corning Incorporated Life Sciences, Tewksbury, MA, USA), which were coated with Matrigel (BD Biosciences), while 800 µl DMEM medium containing 30% FBS was added to the lower chamber. Following culture for 24 h, the uninvaded cells on the upper side of the microporous membrane were removed using a cotton swab. The remaining cells were fixed with 4% paraformaldehyde and stained with 0.5% crystal violet staining solution (Amresco, Solon, OH, USA). The numbers of invaded cells were recorded under an inverted microscope (AE31; Motic Electric) in five randomly-selected fields (magnification, ×200), to obtain the average.
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