Fig. 2
From: Development of syngeneic murine cell lines for use in immunocompetent orthotopic lung cancer models
Validation of a KrasG12D.Smad4+/– cell line. a H&E stain showing orthotopic lung tumor formation by cell line X577. Scale bar is 100 µm and the red line denotes the tumor border. b PCR showing the recombinant KrasG12D allele (305 bp) and Smad4 allele (500 bp) in X577 cells but not in CMT or LLC controls. Although both CMT and LLC cells harbor Kras mutations [16], the 265 bp band in the Kras PCR from CMT and LLC cells represents the wild type (non-engineered) Kras allele. c Western blot showing KRASG12D expression and SMAD4 loss in X577 cells. The KRASG12D-specific antibody detects the KRASG12V mutation in CMT cells but not the KRASG12C mutation in LLC cells. The complete absence of SMAD4 expression suggests that the wild type Smad4 allele has undergone mutation, loss of heterozygosity, or transcriptional silencing