Fig 2: Western blotting detection of VTN and COL1A1 between VUR and non-VUR groups. (A) Western blotting results from similar loaded amounts of proteins stained by Coomassie G250 methods. (B) Western blotting results of VTN and COL1A1 when similar amounts of proteins were loaded between VUR and non-VUR groups. (C) VTN/total protein intensity ratio between VUR and non-VUR groups. (D) COL1A1/total protein intensity ratio between VUR and non-VUR groups. n=10 for each group. ***P<0.001, *P<0.05. VUR, vesicoureteral reflux; COL1A1, α-1 type I collagen; VTN, vitronectin.

Fig 3: Lnc-408 promotes BC cell invasion via LIMK1.A, B qRT-PCR was used to detect Lnc-408 (A) or LIMK1 (B) expression in engineered Hs578T cells (Hs578T/sh Lnc-408, Hs578T/sh Lnc-408/LIMK1 and Hs578T/control) and engineered MCF-7 cells (MCF-7/Lnc-408, MCF-7/Lnc-408/sh LIMK1). C The protein expression levels of LIMK1, p-CREB, CREB, MMP2, ITGB1, and COL1A1 in the indicated BC cells were evaluated by western blotting (band intensity was quantified using GAPDH as a normalizer). D IF staining of p-CREB in the engineered Hs578T and MCF-7 cells described above, captured using fluorescence microscopy (p-CREB was stained red, and nuclei were stained blue with DAPI. Scale bars, 50 μm). E, F Transwell assays to assess the migration and invasion potential of the engineered Hs578T (E) and MCF-7 (F) cells described above (Scale bars, 100 μm). (*P < 0.05, **P < 0.01, ***P < 0.001).

Fig 4: PA-Exo promotes LX-2 cell activation through miR-107-mediated upregulation of IL-9 expression in CD4+ T cells.CD4+ T cells were cocultured with hepatocytes-derived exosomes (Vehicle-Exo, PA-Exo, PA + NCI-Exo, and PA + miR-107I-Exo). Then the culture media were collected and added into culture media of LX-2 cells. CCK-8 and western blot were carried out to evaluate cell viability (A) and the protein levels of α-SMA and CoL1a1 (B) in LX-2 cells. CD4+ T cells were transfected with IL-9 siRNA or scramble control, followed by PA-Exo treatment. CCK-8 and western blot were carried out to evaluate cell viability (C), the protein levels of α-SMA and CoL1a1 (D), and the protein levels of Raf, p-Raf, MEK, p-MEK, ERK, and p-ERK (E) in LX-2 cells. ****P < 0.0001 vs. Vehicle-Exo; ###P < 0.001, ####P < 0.0001 vs. PA + NCI-Exo; $$$P < 0.001, $$$$P < 0.0001 vs. Scramble; &P < 0.05, &&P < 0.01, &&&P < 0.001 vs. IL-9 siRNA. Data were expressed as the mean ± standard deviation (n = 3).

Fig 5: PA-Exo induces LX-2 cell activation by transferring miR-107 to LX-2 cells.A QRT-PCR was performed to detect miR-107 expression in Vehicle-Exo and PA-Exo. B QRT-PCR was performed to determine the transfection efficiency of miR-107I in hepatocytes. The hepatocytes-derived exosomes were divided into Vehicle-Exo, PA-Exo, PA + NCI-Exo, and PA + miR-107I-Exo, then LX-2 cells were cocultured with them for 16 h, respectively. C The hepatocytes were labeled with the lipophilic fluorescent dye DiO (green) and co-incubated with LX-2 cells transfected with mCherry plasmid (red). The laser confocal microscope was used to analyze the intake of exosomes by LX-2 cells (Scale bar = 10 μm). QRT-PCR, CCK-8, and western blot were performed to detect miR-107 expression (D), cell viability (E), and the protein levels of α-SMA and CoL1a1 (F) in LX-2 cells co-incubated with Vehicle-Exo, PA-Exo, PA + NCI-Exo, and PA + miR-107I-Exo, respectively. ****P < 0.0001 vs. Vehicle-Exo; ###P < 0.001 vs. miR-107I-Exo; $$$P < 0.001 vs. PA + NCI-Exo; $$$$P < 0.0001 vs. PA + NCI-Exo. Data were expressed as the mean ± standard deviation (n = 3).