Additional file 1: of Restoration of mutant K-Ras repressed miR-199b inhibits K-Ras mutant non-small cell lung cancer progression
Creators: name: Jin, Hua
name: Jang, Yoonjeong
name: Cheng, Nian
name: Li, Qing
name: Cui, Peng-Fei
name: Zhou, Zhi-Wei
name: Jiang, Hu-Lin
name: Cho, Myung-Haing
name: Westover, Kenneth
name: Tan, Qun-You
name: Xu, Cheng-Xiong
Publisher: name: figshare
Year: 2019
Subjects: subject: Biophysics
subject: Biochemistry
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Physiology
subject: Developmental Biology
subject: Cancer

description: Table S1 Primer sequences used for qRT-PCR. Figure S1 Expression of K-Ras in NSCLC cells that overexpressed or silenced K-Ras. a shRNA of K-Ras significantly suppressed K-Ras expression in NSCLC cells. Indicated cells were transfected with indicated shRNA of K-Ras. After 72 hours of transfection, cells were subjected to Western blot analysis. b Transfection of K-Ras (G12D) expression vector significantly increased K-Ras (G12D) expression in NSCLC cells. Indicated cells were transfected with K-Ras (G12D) expression plasmid. After 72 hours of transfection, cells were subjected to Western blot analysis. Figure S2 Expression of miR-199b in NSCLC cells that overexpressed or inhibited miR-199b. a H522 and H1975 cells were transfected with indicated negative oligonucleotides control (NC) or antisense nucleotides of miR-199 (ASO miR-199b). After 72 hours of transfection, cells were subjected to qRT-PCR analysis. b miR-199b was significantly decreased in stably expressing miR-199b antisense H522 cells. c A549 and H2122 cells were transfected with indicated negative oligonucleotides control (NC) or miR-199 mimics. After 72 hours of transfection, cells were subjected to qRT-PCR analysis. d miR-199b level was significantly increased in stably expressing miR-199b A549cells. Figure S3 Aerosol delivery of miR-199b to the lung of mice. a Gene delivery efficiency of PCAmHn as a gene carrier. Delivery efficiency of PCAmHn as a gene carrier was evaluated using PCAmHn/green fluorescent protein (GFP) expression plasmid complex. ICR mice were exposed to aerosol containing PCAmHn/GFP expression plasmid complex or GFP expression plasmid only for 30 minuets, and 72 hours post-treatment, the mice were sacrificed for delivery efficiency assay. Green signals indicated that most of the delivered GFP was efficiently trasfected into lung. b miR-199b expression was measured in the lung tissue of K-RasLA1 transgenic mice. Control mice were exposed to the gene carrier only (carrier); Vector group mice were exposed to vector mixed with the gene carrier (vector); miR-199b group mice were exposed to the miR-199b expression plasmid mixed with the gene carrier (miR-199b). Figure S4 The protein levels of miR-199b candidate targets in NSCLC cells and the lungs of K-RasLA1 transgenic mice. a. The expression levels of the indicated proteins in Fig. 4e were quantified using image J software. b. The expression levels of the indicated proteins in Fig. 4f were quantified using image J software. *,p&lt;0.05 compare to carrier control; **, p&lt;0.01 compared to carrier control; #, p&lt;0.05 compared to vector control; ##, p&lt;0.01 compared to carrier control. Figure S5 The protein levels of Akt and ERK signaling pathway related genes in NSCLC cells. a. The expression levels of the indicated proteins in Fig. 5a were quantified using image J software. b. The expression levels of the indicated proteins in Fig. 5c were quantified using image J software. (PPTX 820 kb)