Fig 1: Characterization of treatment efficacy.a Schematic of cytosolic drug delivery through membrane fusion for treatment in vitro and in vivo. b Gene silencing ability (left) and protein knock-down effect (right) of siR-Sox2 loaded CNVs, eT-CNVs, and eFT-CNVs in HepG2 cells (n = 6; p < 0.05, one-way ANOVA). The equivalent concentration of siRNA was 200 nM. c Quantitative data of cell cycles of 2 × 105 HepG2 treated with 200 nM free paclitaxel or paclitaxel equivalent in nanovesicles for 48 h, respectively (n = 3; p < 0.05, one-way ANOVA). d IC50 of siR-Sox2 (left, n = 6), gelonin (middle, n = 6), and paclitaxel (right, n = 6) loaded three types of nanovesicles treated HepG2 cells, respectively. e Quantitative data of wound closure showing free drugs and drugs loaded nanovesicles inhibited migration of HepG2 cells compared to the negative control (n = 12; p < 0.001, one-way ANOVA). f Quantitative data of cell proliferation of HepG2 treated with drugs and drugs-loaded nanovesicles for 48 h, respectively (n = 200; p < 0.05, one-way ANOVA). g Quantitative data of the invasion assay showing free drugs and drugs loaded nanovesicles inhibited the invasion of HepG2 cells compared to the negative control (n = 6; p < 0.05, one-way ANOVA). h Quantitative data of tumor volume of HepG2 cells xenograft in mice from siR-Sox2 (left, n = 5; p < 0.001, one-way ANOVA), gelonin (middle, n = 5; p < 0.001, one-way ANOVA), and paclitaxel (right, n = 5; p < 0.001, one-way ANOVA) loaded group after drug or placebo administration. i Biodistribution of paclitaxel in tumors in free paclitaxel and paclitaxel loaded eFT-CNV groups, respectively, after an intravenous administration (n = 6; p < 0.001, one-way ANOVA).
Fig 2: ATRA treatment reduces the expression of stemness markers SOX2 and NES in GBM cell lines. Relative mRNA expression levels of (A) SOX2 and (B) NES in U87-MG cells, and (C) SOX2 and (D) NES in A172 cells, following treatment with vehicle (DMSO) or 1 µM ATRA for 5 days. Expression was quantified by qPCR, normalized to the geometric mean of GAPDH and ACTB, and calculated relative to the vehicle control group (set to 1) using the ΔΔCt method. Data are presented as mean fold change ± SEM from three biological replicates. Statistical significance was determined by Student’s t-test. ** p < 0.01, *** p < 0.001.
Fig 3: U87-MG and A172 protein validation. Consistent with the mRNA findings, ELISA quantification demonstrated robust reductions at the protein level (Figure 2). In U87-MG, SOX2 protein declined from 148.33 ± 6.01 to 49.67 ± 2.73 pg µg−1 (0.335-fold, p = 0.0002) and Nestin from 450.00 ± 17.32 to 128.33 ± 4.41 pg µg−1 (0.285-fold, p = 0.00008). A172 showed analogous decreases—SOX2: 123.33 ± 5.87 to 53.33 ± 2.40 pg µg−1 (0.432-fold, p = 0.0007); Nestin: 403.33 ± 14.53 to 144.33 ± 3.53 pg µg−1 (0.358-fold, p = 0.00009). ** p < 0.01, *** p < 0.001.
Supplier Page from Abcam for Human SOX2 ELISA Kit