Supplementary Figure S2 from A Sequential Targeting Strategy Interrupts AKT-Driven Subclone-Mediated Progression in Glioblastoma
openalex（2023）
Abstract
Extended Data relating to Main Figure 2/Assessment of ALDH1A1+ cells from primary, treatment-naive glioblastoma. A, Bar plot shows ALDH-bright (ALDHbr) vital cells from two additional paired cases of naive vs. experimental (TMZ→eR) and clinical (cR) relapse. Data as mean ± SD. p values by one-way analysis of variance (ANOVA) with Tukey´s post-hoc test. B, Dotplot shows ALDH1A1 mean fluorescence intensity (MFI) in paired BN46 treatment-naive vs. experimental relapse conditions (in vitro exposure to TMZ (TMZ→eR) or irradiation (RT→eR)). Data are normalized to isotype control, mean ± SD. p values calculated by Kruskal-Wallis test with Dunn's post-hoc test. C, Relative ALDH1A1 expression in -knockdown (shALDH1A1) and -overexpression (Ovx) BN46 cells used for indicated experiment. Data shown as mean ± SD, normalized to their respective controls (ALDH1A1 Ovx to GFP Ctrl | shALDH1A1 to shNT Ctrl). D, Left: Brightfield image of BN46 cells in 96-well plates during monitoring by software-based cell recognition in the limiting dilution assay (NyOne®). An exemplary single cell/well is shown at one day post seeding; representative monoclonal colonies of ALDH1A1-knockdown (sh) and -overexpressing (Ovx) cells at day 16 after seeding. Right: Doubling time estimated at day 16 after seeding (see Methods). Data as mean ± SD, p values calculated by Kruskal-Wallis test with Dunn's post-hoc test. E, Left: Cartoon describes the neurosphere experiments shown in Fig. 2H. Right: Phase contrast microscopic appearance of plated 2° neurosphere and respective immunofluorescence visualization of antibody labeling on neurosphere-derived cells. Neuronal phenotype, β3-tubulin (β3-tub); glial phenotypes, glial fibrillary acidic protein (GFAP). Nuclei exposed with DAPI. Scale bars: left: 100 µM, right: 50 µM. F, Table showing quantification data of 1° and 2° neurosphere generations from (E) in the respective treatment-naive BN46 cells. G, All plated 2° neurospheres from assay (E,F) generated neuronal and glial cell phenotypes.
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