Journal: International Journal of Molecular Sciences

Article Title: High SOX9 Maintains Glioma Stem Cell Activity through a Regulatory Loop Involving STAT3 and PML

doi: 10.3390/ijms23094511

Figure Lengend Snippet: Genetic and pharmacological STAT3 inhibition in glioma cells: ( A ) representative immunoblots of p-STAT3, STAT3, PML, SOX9, and SOX2 in U251-MG and GNS166 cells in STAT3 knockdown (“ sh41 ” and “ sh43 ”) and control (“ pLKO ”) cells ( n = 3); ( B ) quantification of cell growth in sh41 and sh43 cells compared with controls ( n = 2); ( C ) quantification of SA-β-gal + cells in STAT3 knockdown cells ( n = 2); ( D ) quantification of tumorspheres (primary and secondary) in sh41 and sh43 U251-MG cells relative to pLKO condition ( n = 2); ( E ) tumor volume representation at indicated time points after subcutaneous injection of U251-MG cells ( n = 4); ( F ) ELDA plot of limiting dilution assay of sh41, sh43 , and control U251-MG cells; ( G ) cytotoxicity exhibited by indicated glioma cells after treatment with increasing doses of STX-0119 for 72 h ( n = 6); ( H ) quantification of tumorspheres in indicated cells after treatment with 50 and 100 µM STX-0119 ( n = 3). Calculations were relative to untreated cells; ( I ) representative immunoblot of p-STAT3, STAT3, PML, SOX9, and SOX2 expression in U251-MG cells after treatment with 50 and 100 µM STX-0119 for 72 h ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Cells were seeded in 96-well plates at a density of 2.5 × 10 3 cells per well and treated 24 h later with the indicated concentrations of STX-0119 (Sigma-Aldrich) for 72 h or arsenic trioxide (ATO; Sigma-Aldrich) for 48 h, in sextuplicates.

Techniques: Inhibition, Western Blot, Injection, Limiting Dilution Assay, Expressing