FIGURE 2 from Activation of KrasG12D in Subset of Alveolar Type II Cells Enhances Cellular Plasticity in Lung Adenocarcinoma
Year: 2023
Type: other
Abstract: Double-positive cells in Sftpc-CreER; LSL-KrasG12D-induced lung adenocarcinoma are tumor-propagating cells and forms tumors upon transplantation. A, Experimental outline for growing cells in 3D organoid culture. B, FACS scatter plot showing single (type I or type II) and double-positive population sorted from Sftpc-CreER; LSL-KrasG12D; TdTomato mice. Type I (TdTm−Pdpn+), type II (TdTm+ Pdpn−), and double-positive (type I/II+; TdTm+Pdpn+) fractions were sorted. C, Brightfield images of 3D organoid cultures from different indicated sorted populations on day 14. Colony number (D) and colony size (E) was quantified using ImageJ software. F, 3D organoid culture from double-positive cells on day 14 in red channel. G, Immunofluorescent image on day 14 of organoid culture showing double-positive cells differentiate into type I and type II cells. Ki67 (H) and TTF1 (I) staining of 3D organoid culture. J, Representative H&E staining of tumors sections which were obtained after subcutaneous transplantation of double-positive (type I/II+) cells. Right panel shows higher magnification of boxed area in left panels. K, Representative IHC images of tumor sections for TTF1. Right panel shows higher magnification of boxed area in left panels. L, Representative immunofluorescent image for in red channel showing Tdtomato expression in tumor obtained after transplantation. M, Representative IF images for Sftpc and Rage staining. Right panel shows higher magnification of boxed area in left panels. Results shown in B, F, G, H, I, J, K, L, and M are representative of three independent experiments. D and E represent data from three independent experiments. Error bars represent the mean ± SEM values and significance is defined as *, P ≤ 0.05; **, P ≤ 0.01; and ***, P ≤ 0.001.
Language: English
Topic: Cancer Cells and Metastasis
Subfield: Oncology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W4388970058