Supplementary Figure S2 from A Sequential Targeting Strategy Interrupts AKT-Driven Subclone-Mediated Progression in Glioblastoma Article Swipe

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Sied Kebir , Vivien Ullrich , Pia Berger , Celia Dobersalske , Sarah Langer , Laurèl Rauschenbach , Daniel Trageser , Andreas Till , Franziska K. Lorbeer , Anja Wieland , Timo Wilhelm-Buchstab , Ashar Ahmad , Holger Fröhlich , Igor Cima , Shruthi Prasad , Johann Matschke , Verena Jendrossek , Marc Remke , Barbara M. Grüner , Alexander Roesch , Jens T Siveke , Christel Herold-Mende , Tobias Blau , Kathy Keyvani , Frank K.H. van Landeghem , Torsten Pietsch , Jörg Felsberg , Guido Reifenberger , Michael Weller , Ulrich Sure , Oliver Brüstle , Matthias Simon , Martin Glas , Björn Scheffler ·

YOU? · · 2025 · Open Access · · DOI: https://doi.org/10.1158/1078-0432.30706284

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.

Related Topics

Glioblastoma

Immunofluorescence

Glial Fibrillary Acidic Protein

Phase-Contrast Microscopy

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Neurosphere Glioblastoma Immunofluorescence Glial fibrillary acidic protein Monoclonal antibody Isotype Pathology Contrast (vision) Antibody Immunohistochemistry Phase contrast microscopy Biology In vitro Coefficient of variation Chemistry Cell culture Nuclear medicine Molecular biology Limiting Doubling time Intensity (physics)

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Title: Supplementary Figure S2 from A Sequential Targeting Strategy Interrupts AKT-Driven Subclone-Mediated Progression in Glioblastoma

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Authors: Sied Kebir, Vivien Ullrich, Pia Berger, Celia Dobersalske, Sarah Langer, Laurèl Rauschenbach, Daniel Trageser, Andreas Till, Franziska K. Lorbeer, Anja Wieland, Timo Wilhelm-Buchstab, Ashar Ahmad, Holger Fröhlich, Igor Cima, Shruthi Prasad, Johann Matschke, Verena Jendrossek, Marc Remke, Barbara M. Grüner, Alexander Roesch, Jens T Siveke, Christel Herold-Mende, Tobias Blau, Kathy Keyvani, Frank K.H. van Landeghem, Torsten Pietsch, Jörg Felsberg, Guido Reifenberger, Michael Weller, Ulrich Sure, Oliver Brüstle, Matthias Simon, Martin Glas, Björn Scheffler

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Concepts: Neurosphere, Glioblastoma, Immunofluorescence, Glial fibrillary acidic protein, Monoclonal antibody, Isotype, Pathology, Contrast (vision), Antibody, Immunohistochemistry, Phase contrast microscopy, Biology, In vitro, Coefficient of variation, Chemistry, Cell culture, Nuclear medicine, Molecular biology, Limiting, Doubling time, Intensity (physics)

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publication_date	2025-11-25
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