Fig. 2: Gal-3 is required for macropinocytosis.
From: Macropinocytosis requires Gal-3 in a subset of patient-derived glioblastoma stem cells
a Immunoblots show expression of indicated proteins for Ge518, Ge269, and Ge835 infected by shRNA Control (Ctrl) or shGal-3. Histograms show the fold change of protein expression determined by densitometry analysis. b Macropinocytosis uptake assay using TMR-dextran in shCtrl vs. shGal-3 #2 GSCs. Scale bar, 10 µm. Histograms represent the fold change of macropinocytosis activity in Ge518, Ge269, and Ge835 normalized to nuclei number (n = 4–6). Ctrl = Vehicle (DMSO). c Cell viability of Ge518, Ge269, and Ge835 in shRNA Control (Ctrl) vs. shGal-3, measured by CellTiter-Glo in GSCs (n = 3–4). d Cell viability of Ge518 shRNA Control (Ctrl) vs. shGal-3 under EIPA treatment, measured by CellTiter-Glo in Ge518 (n = 2–3). Data are represented as mean ± SEM (*p < 0.05, **p < 0.01, and ***p < 0.001), two-way ANOVA, Dunnett’s multiple comparisons test. e Cell invasion in 3D of Ge518 shRNA Control (Ctrl) vs. shGal-3 under EIPA treatment (n = 2–3). Scale bar, 100 µm. Histograms represent the fold change of the invasion score in Ge518. Data are represented as mean ± SEM (*p < 0.05, **p < 0.01, and ***p < 0.001), two-way ANOVA, Dunnett’s multiple comparisons test. f Effect of Gal-3 knockdown on tumor growth in vivo: Ge269 shCtrl vs. shGal-3, (n = 7 mice per group), p = 0.016 (Log-rank Mantel–Cox test). g Histological analysis of Ge269 shCtrl vs. shGal-3. Tumors were stained for Gal-3, and counterstained with hematoxylin. (n = at least 3 mice per group). Scale bar, 50 µm. Data are represented as mean ± SEM (*p < 0.05, **p < 0.01, and ***p < 0.001), two-way ANOVA, Sidak’s adjusted p value. ns = nonsignificant, Ctrl = Vehicle (DMSO).
