Figure 1 from Membrane-bound Heat Shock Protein mHsp70 Is Required for Migration and Invasion of Brain Tumors
Authors: Shevtsov Maxim , Bobkov Danila , Yudintceva Natalia , Likhomanova Ruslana , Kim Alexander , Fedorov Evegeniy , Fedorov Viacheslav , Mikhailova Natalia , Oganesyan Elena , Shabelnikov Sergey , Rozanov Oleg , Garaev Timur , Aksenov Nikolay , Shatrova Alla , Ten Artem , Nechaeva Anastasiya , Goncharova Daria , Ziganshin Rustam , Lukacheva Anastasiya , Sitovskaya Daria , Ulitin Alexey , Pitkin Emil , Samochernykh Konstantin , Shlyakhto Evgeny , Combs Stephanie E. 
Published: August 2024

<p>Evaluation of the membrane-bound Hsp70 expression in malignant brain tumors by live-cell confocal microscopy imaging. <b>A,</b> Schematic description of the study of brain tumor samples to determine the expression of membrane-associated chaperone in cancer cells, followed by mass spectrometry analysis of isolated lipid rafts. <b>B,</b> Representative confocal microscopy images of the brain tumor samples stained for Hsp70 (green), TMRM (tetramethylrodamine methyl ester; red), disialoganglioside GD2 (red), and protoporphyrin IX (pPIX; magenta). Nuclei were stained by DAPI. Scale bars, 100 μm. Respective MR images (postcontrast T1-weighted and FLAIR images) of the patients are presented. <b>C,</b> Representative confocal microscopy images of the brain metastasis stained for Hsp70 (green), TMRM (tetramethylrodamine methyl ester; red). Nuclei were stained by DAPI (blue). Scale bars, 100 μm. Respective MR images (postcontrast T1-weighted and FLAIR images) of the patients as well as hematoxylin and eosin staining of tumor samples are presented. <b>D,</b> Top row—Fluorescence intensity measurements of mHsp70 expression in live-cell microscopy images of patients with GBM (<i>n</i> = 11) with subsequent Benjamini–Yekutieli test for multiple pairwise comparison of protein expression levels. The bar graph shows the median ± 95% CI (Man–Whitney test result—****, <i>P</i> < 0.0001). The heat map shows <i>P</i> values as the result of a <i>post hoc</i> multiple comparison test. Bottom row—Fluorescence intensity measurements of mHsp70 expression in live-cell microscopy images of brain metastasis (<i>n</i> = 3) with subsequent Benjamini–Yekutieli test for multiple pairwise comparisons of protein expression levels. The bar graph shows the median ± 95% CI (Man–Whitney test result—****, <i>P</i> < 0.0001). The heat map shows <i>P</i> values as the result of a <i>post hoc</i> multiple comparison test. <b>E,</b> Representative multiplex analysis of the immunofluorescence of brain tumor sample stained for Hsp70, SOX2, and nestin. Nuclei were stained by DAPI. NA, necrotic area; VT, vital tissues. Scale bars, 200 μm. <b>F,</b> Representative confocal microscopy images of the normal brain tissue samples stained for Hsp70 (green) and TMRM (tetramethylrodamine methyl ester; red). Nuclei were stained by DAPI (blue). Scale bars, 100 μm. Respective MR images of the patients are presented. <b>G,</b> Live-confocal microscopy images of mHsp70-positive tumor cells in oncostreams detected in GBM samples. Tumor sections were stained for Hsp70 (green), TMRM (red), and DAPI (blue) for nuclei. Scale bars, 100 μm. For the descriptive characteristics of patients, see Supplementary Table S1.</p>