Fig. 1: EGFR R252C promotes tumor cell proliferation and tumor growth in vivo.
From: An alternative EGFR activation by patient-derived R252C mutation promotes cancer progression
a Chest CT and brain MRI images of a patient with multifocal lung cancer and glioma. b Treatment timeline of the patient. c DNA sequencing of genomic DNA from U87, U251, or H1299 cells with or without EGFR R252C knock-in. d, e Immunoblotting assay was performed with U87, U251, and H1299 cells with or without EGFR R252C knock-in. Tubulin was used to determine equivalent loading. IB, immunoblotting. d Representative images of immunoblotting were shown. e Semi-quantitative scoring was carried out. The EGFR/Tubulin ratio was quantified and normalized to the EGFR WT cells. Rel, relative. f U87, U251 or H1299 cells harboring EGFR WT or R252C mutation were seeded in 96-well plates. Cell proliferation assay was performed. Relative cell proliferation of U87, U251, and H1299 cells expressing EGFR WT or R252C was normalized to the day 1 value. g Colony formation assay in U87, U251 or H1299 cells with or without EGFR R252C mutation. A representative experiment is shown in the left panel. The number of colonies was calculated (right panel). h Luciferase-expressing U87 cells with or without EGFR R252C knock-in were intracranially injected into randomized athymic nude mice (five mice per group). 49 days after implantation, bioluminescence imaging of these mice was carried out to examine tumor growth. Data represent the mean ± s.d. of five mice. i Luciferase-expressing H1299 cells with or without EGFR R252C knock-in were injected into the left lung of randomized athymic nude mice (five mice per group). 35 days after implantation, bioluminescence imaging of mice was carried out to examine tumor growth. Data represent the mean ± s.d. of five mice. e, f, g Data represent the mean ± s.d. of three biologically independent experiments. e–i, Unpaired, two-tailed t-test.
