Additional file 2 of Chitinase-3-like protein 1 depletion in glioma cells alters tumor microenvironment and normalizes neovasculature in human glioma xenografts

Supplementary Material 2: Figure 2. a. Gating strategy for the flow cytometric sort of U87-MG-RFP transfected with pCMV-Cas9-GFP plasmid. b-c. Validation of CHI3L1 knock-out in selected candidate clones. b. RT-qPCR analysis of CHI3L1 expression using 2 sets of primers. CHI3L1 expression given as % of WT control. c. ELISA of cell culture supernatants of sgCHI3L1 clones. CHI3L1 concentration given as % of WT control. Red lines indicate the level in WT reference. d. BrdU incorporation assay for U87-MG-RFP cells transfected with control non-targeting gRNA (sgCTR). CRISPR/Cas9 engineering pipeline does not change the proliferation rate of U87-MG-RFP clones. e. Analysis of CHI3L1 expression in NHA, U251, U87-MG and WG9 by RT-qPCR