Journal: International Journal of Molecular Sciences
Article Title: MET Exon 14 Skipping and Novel Actionable Variants: Diagnostic and Therapeutic Implications in Latin American Non-Small-Cell Lung Cancer Patients
doi: 10.3390/ijms252413715
Figure Lengend Snippet: The VUSs predicted to be drivers, T992I and H1094Y, promote the survival of proliferative non-tumor cells and migration in tumor cells. ( A ) Representative Western blots of total Met and β-actin expression, evaluated for the H1993 GFP (basal), METex14, T992I, and H1094Y cells. ( B ) Densitometry levels of total normalized Met/β-actin (+SEM). The graph represents the normalized average from 3 independent experiments, ±SEM. ( C ) The representative Western blots of total Met and β-actin expression were evaluated for the HEK293T GFP (basal), METex14, T992I, and H1094Y cells. ( D ) Densitometry levels of total normalized Met/β-actin (+SEM). The graph represents the normalized average from 3 independent experiments, ±SEM. ( E ) The absorbance averages of HEK293T and H1993 cells expressing GFP, METex14, T992I, and H1094Y; the cells incubated with and without HGF. ( F ) Representative microphotographies of the wound healing at 0 and 24 h of H1993 cells expressing METex14, T992I, and H1094Y, treated with and without HGF were taken at 4×. ( G ) The wound closure percentage was calculated for each experimental condition. Finally, three independent experiments averaging the ±SEM are shown. A two-way ANOVA with Tukey correction was applied, and the p -values were adjusted for multiple comparisons. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; n.s. non-significant.
Article Snippet: METex14 (pLV[Exp]-Puro-CMV (hMET[NM_001127500.3]*(delete exon 14)) (VB210712-1387jhm), T992I (pLV[Exp]-Puro-CMV (hMET[NM_000245.4]*(T992I)) (VB210720-1190man), and H1094Y (pLV[Exp]-Puro-CMV (hMET[NM_000245.4]*(H1094Y)) (VB210720-1189kqp) were designed using Vector Builder (Chicago, IL, USA), and the pLentiCMV MET GFP Puro (Addgene #37560, Watertown, MA, USA) was acquired in Addgene.
Techniques: Migration, Western Blot, Expressing, Incubation
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