Fig 1: Overexpression of circENTPD7 decreased the levels of PTEN protein in NSCLC cells. To explore the effects of circENTPD7 on PTEN, NCI-H1703 and HCC4006 cells were transfected with circENTPD7 expression vector (A). The effects of circENTPD7 on PTEN mRNA (B) and PTEN protein (C) were determined by RT-qPCR and Western blot, respectively. Mean ± SD values were used to express data of three biological replicates. *, p < 0.05.
Fig 2: PTEN was downregulated in NSCLC and was inversely correlated with circENTPD7. Expression of PTEN mRNA (A) and protein (B) in the 64 pairs of tissues were determined by RT-qPCR and ELISA, respectively. Average values of three technical replicates were calculated to express the data of gene expression in paired tissues. ***, p < 0.001. Pearson’s correlation was carried out to explore the correlations between circENTPD7 and PTEN mRNA (C) or PTEN protein (D).
Fig 3: Anti-inflammatory effects of AdMSCs-derived exosomes via ROCK1/PTEN pathway. (a) The expression levels of ROCK1 and PTEN were quantified by real time-qPCR after treatment with ROCK1 and PTEN inhibitor. (b) THP-1 cells were treated with 100 ng/mL LPS (L), LPS + 5 µg/mL exosomes (LE), LPS + exosomes + 10 nM ROCK1 inhibitor (LE + Ri), and LPS + exosomes + 1 nM PTEN inhibitor (LE + Pi). (c) Protein levels of ROCK1 and PTEN were analyzed by western blot. ß-actin was used as controls. Relative protein levels were confirmed by quantification. (d) A schematic diagram of miR-124-3p binding sites in the 3’ UTR of ROCK1 mRNA. After treatment with exosomes, the relative level of miR-124-3p was determined in THP-1 cells by real time-qPCR. The data are expressed as the mean ± SD of three independent experiments. *Significant difference from untreated control (p < 0.05). **Significant difference from untreated control (p < 0.01).
Fig 4: Overexpression of circENTPD7 increased NSCLC cell proliferation through PTEN. The role of circENTPD7 and PTEN in regulating the proliferation of NSCLC cells was explored by CCK-8 assay. Mean ± SD values were used to express data of three biological replicates. *, p < 0.05.
Fig 5: Effect of AdMSCs-derived exosomes on the inflammatory response. (a) THP-1 cells were treated with 100 ng/mL LPS and LPS + 5 µg/mL exosomes for 24 h. Untreated cells were used as controls. Real time-qPCR was performed to investigate the changes of relative gene expressions on inflammatory- (TNF-a, IL-6, IL-8) and anti-inflammatory mRNAs (CD163, Arg1, CD206, TGF-ß1, IL-10). (b) The expression of ROCK1 and PTEN was analyzed in THP-1 cells after treatment with LPS and LPS + exosomes for 24 h by real time-qPCR. GAPDH was utilized as an internal control. (c) The protein levels of ROCK1 and PTEN were confirmed by Western blotting. ß-actin was used as an internal control. The relative values of ROCK1/ß-actin and PTEN/ß-actin were determined by Image J program. The data are expressed as the mean ± SD of three independent experiments. *Significant difference from untreated control (p < 0.05). **Significant difference from untreated control (p < 0.01).
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