Fig 1: Visualizing neural crest lineage cells in Tsc2+/- mouse renal tumors(A–D) Representative tdTomato and merged tdTomato/DAPI images of immunohistochemical slices (4 µM thickness) of renal tumors showing punctate singlets and colonies of tdTomato+ neural crest cell populations obtained in 6-month-old (A and B) and 12-month-old (C and D) Tsc2+/-Mpz(Cre)TdTfl/fl mice viewed using an excitation filter of 540/45 nm in a confocal microscope.(E–H) (E) Comparative image panel of renal tissue from wild-type 12-month-old mice. Double immunolabeling of renal tumor slices obtained from 16-month-old Tsc2+/-Mpz(Cre)TdTfl/fl mice reveal tandem endogenous expression of Mpz-driven tdTomato red fluorescent protein (F), Alexa Fluor 488-labeled CD57 neural crest marker expression (G), and tdTomato/CD57/DAPI merged images (H) confirming neural crest identity of these cells. Confocal and fluorescence imaging was performed using N = 3 slides; 2 sections/slide to visualize each renal tumor/tissue. Scale bar represents 50µm. See also Figures S1D and S1E for fluorescent imaging of immunohistochemical slices of hepatic lesions of Tsc2+/-Mpz(Cre)/TdTfl/fl mice showing punctate tdTomato+ NCCs.