Journal: Cells

Article Title: Atypical Exon 2/3 Mutants G48C, Q43K, and E37K Present Oncogenic Phenotypes Distinct from Characterized NRAS Variants

doi: 10.3390/cells13201691

Figure Lengend Snippet: Effect of the novel NRAS mutants on cell proliferation. NRAS G48C, Q43K, and E37K enhanced proliferation of both HCT116 and NIH3T3 cells compared to vector-only, wild-type, and canonical mutant control Q61K. Proliferation rates of ( a ) HCT116 cells and ( b ) NIH3T3 cells transfected with empty vector, wild-type NRAS, canonical NRAS Q61K mutant, and the novel NRAS mutants G48C, Q43K, and E37K are shown. Data presented are representative of three independent trials, each performed in triplicate, and expressed as mean ± standard deviation (SD). * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001. PTT = pTargeT vector-only control; WT = wild-type; ns = non-significant.

Article Snippet: Two cell lines were used in this study, as follows: the NIH3T3 murine embryonic fibroblast cell line (Cat. No. CCL-247) and the HCT116 human colorectal carcinoma cell line (Cat. No. CCL-247), which were both obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Plasmid Preparation, Mutagenesis, Control, Transfection, Standard Deviation

Journal: Cells

Article Title: Atypical Exon 2/3 Mutants G48C, Q43K, and E37K Present Oncogenic Phenotypes Distinct from Characterized NRAS Variants

doi: 10.3390/cells13201691

Figure Lengend Snippet: Effect of the novel NRAS mutants, G48C, Q43K, and E37K, on cell survival. ( a ) HCT116 and ( b ) NIH3T3 cells were transfected with NRAS WT and mutant constructs, then assessed for their ability to resist apoptosis when induced with sodium butyrate (HCT116) and reduced serum concentration (NIH3T3). Data presented are representative of three independent trials, each performed in triplicate, and expressed as mean ± standard deviation (SD). * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001. PTT = pTargeT vector-only control; WT = wild-type; ns = non-significant.

Article Snippet: Two cell lines were used in this study, as follows: the NIH3T3 murine embryonic fibroblast cell line (Cat. No. CCL-247) and the HCT116 human colorectal carcinoma cell line (Cat. No. CCL-247), which were both obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Transfection, Mutagenesis, Construct, Concentration Assay, Standard Deviation, Plasmid Preparation, Control

Journal: Cells

Article Title: Atypical Exon 2/3 Mutants G48C, Q43K, and E37K Present Oncogenic Phenotypes Distinct from Characterized NRAS Variants

doi: 10.3390/cells13201691

Figure Lengend Snippet: Effect of NRAS G48C, Q43K, and E37K mutants on cellular migration in HCT116 and NIH3T3 cells. Scratch wound assay showing representative images of ( a ) HCT116 and ( c ) NIH3T3 cells transfected with empty vector, wild-type NRAS, canonical NRAS Q61K control, and the novel NRAS mutants G48C, Q43K, and E37K at 0 h and 16 h post-scratch. The migration rates for each setup are shown in ( b , d ) for HCT116 and NIH3T3 cells, respectively. Data presented are representative of three independent trials, each performed in triplicate, and expressed as mean ± standard deviation (SD). ** p ≤ 0.01. pTT = pTargeT vector-only control; WT = wild-type; ns = non-significant.

Article Snippet: Two cell lines were used in this study, as follows: the NIH3T3 murine embryonic fibroblast cell line (Cat. No. CCL-247) and the HCT116 human colorectal carcinoma cell line (Cat. No. CCL-247), which were both obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Migration, Scratch Wound Assay Assay, Transfection, Plasmid Preparation, Control, Standard Deviation

Journal: Cells

Article Title: Atypical Exon 2/3 Mutants G48C, Q43K, and E37K Present Oncogenic Phenotypes Distinct from Characterized NRAS Variants

doi: 10.3390/cells13201691

Figure Lengend Snippet: Expression of NRAS mutants induce cytoskeletal remodeling in NIH3T3 cells. Distinct features, such as prominent peripheral stress fibers (yellow arrows) and highly organized actin filaments (red arrowheads), are present in both the empty vector ( A ) and wild-type ( B ) setups. Central round nuclei (orange arrowheads) are also visible in WT. Changes in cytoskeletal architecture, as well as invasive structures, are observable in both canonical and novel mutant setups ( C , D = Q61K; E , F = G48C; G , H = Q43K; I , J = E37K), including fan-shaped cells with prominent migrating front (sky blue brackets); peripheral dorsal ruffles (pink arrowheads); circular dorsal ruffles (yellow arrowheads); tunneling nanotubes (gray arrowheads); multilobulated nuclei (red arrows); invadopodia (pink arrows); lamellipodia (gold arrowheads); filopodia (purple arrowheads); and fusing cells (yellow brackets).

Article Snippet: Two cell lines were used in this study, as follows: the NIH3T3 murine embryonic fibroblast cell line (Cat. No. CCL-247) and the HCT116 human colorectal carcinoma cell line (Cat. No. CCL-247), which were both obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Expressing, Plasmid Preparation, Mutagenesis