Additional file 1 of Chloride intracellular channel 1 activity is not required for glioblastoma development but its inhibition dictates glioma stem cell responsivity to novel biguanide derivatives
Creators: name: Barbieri, Federica
name: Bosio, Alessia Graziana
name: Pattarozzi, Alessandra
name: Tonelli, Michele
name: Bajetto, Adriana
name: Verduci, Ivan
name: Cianci, Francesca
name: Cannavale, Gaetano
name: Palloni, Luca M. G.
name: Francesconi, Valeria
name: Thellung, Stefano
name: Fiaschi, Pietro
name: Mazzetti, Samanta
name: Schenone, Silvia
name: Balboni, Beatrice
name: Girotto, Stefania
name: Malatesta, Paolo
name: Daga, Antonio
name: Zona, Gianluigi
name: Mazzanti, Michele
name: Florio, Tullio
Publisher: name: figshare
Year: 2022
Subjects: subject: Biochemistry
subject: Medicine
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Physiology
subject: Pharmacology
subject: Biotechnology
subject: 39999 Chemical Sciences not elsewhere classified
subject: FOS: Chemical sciences
subject: Science Policy
subject: Developmental Biology
subject: Cancer
subject: 110309 Infectious Diseases
subject: FOS: Health sciences

description: Additional file 1: Figure S1. Immunohistochemical distribution of CLIC1 in GBM. Figure S2. A) CLIC1 expression in GSCs isolated from 14 human GBMs, evaluated by RNA-seq, and expressed as counts per million reads mapped (CPM). Only GBM 39 (arrow) displayed low CLIC1 mRNA content. The Table reports the corresponding codes of the GSC cultures used in this study and the RNA-seq data deposited at NCBI Geo data set. B) Upper panel: CLIC1 expression evaluated by WB, in total cell lysates from selected GSC cultures. Membranes were re-probed with α-tubulin antibody after stripping and used as a reference for protein loading. Lower panel: Histograms report CLIC1/α-tubulin ratio of densitometric values and expressed in arbitrary units (A.U.) as mean ± S.D. Figure S3. Dose-response curves of novel biguanide derivatives and metformin on individual GSC cultures. The average response is reported in the Fig. 1A of the manuscript. Figure S4. Dose-response curves of novel biguanide derivatives and metformin on individual non-stem differentiated GBM cell cultures (GBM D). The average response is reported in the Fig. 1B of the manuscript. Figure S5. Dose-response curves of metformin and novel biguanide derivatives on individual ucMSC cultures. The average response is reported in the Fig. 1C of the manuscript. Figure S6. Kaplan-Mayer curves of Q48, Q54, and metformin (MET) depicting the effect of supramaximal concentration of these compounds on zebrafish embryos survival. Limited toxicity, not different from controls, was observed for all the compounds up to 5 days of treatment. Experiments were repeated twice, n = 20 per experimental group. Q48: log rank test for trend p = 0.62; Q54: log rank test for trend p = 0.64; metformin: log rank test for trend p = 0.38. Figure S7. Effect of different doses of metformin and Q54 on chick embryo survival after 10 days of incubation. Experiments were performed by Inovotion (La Tronche, France). No toxicity was observed for these compounds up to 3 mM. Experiments were repeated twice, n = 18 per experimental group. Figure S8. A. Dose-response curves of Q46, Q48, Q54 and metformin on rat astrocyte cultures. Limited toxicity is observed for all the novel compounds. Only metformin reduced astrocyte viability (− 50%) at the highest concentration tested (30 mM). Data are expressed as average of experiments preformed in quadruplicate and repeated twice. B. Table reports IC50 values in non-malignant rat astrocytes and GSCs and the calculated selectivity indices for each compound. According to the “selectivity criteria” all biguanides are considered selective compounds against GSCs (selectivity index &gt; 10) (see reference [62]). Figure S9. A. CLIC1 expression in GBM19 GSCs carrying siRNA for both Luciferase (siLuc, silencing control) and CLIC1 (siCLIC1). B. Cell proliferation of GBM19 siLuc and GBM19 siCLIC1, treated with Q48 and Q54 (100 μM) for 48 h and evaluated by counting live cells. Data represent the mean ± S.E.M. ***p