Journal: Journal of nanobiotechnology

Article Title: Evaluating the pro-survival potential of apoptotic bodies derived from 2D- and 3D- cultured adipose stem cells in ischaemic flaps.

doi: 10.1186/s12951-024-02533-1

Figure Lengend Snippet: Fig. 6 3D-ABs suppress oxidative stress and cell death, and stimulated angiogenesis in ischaemic flaps. (A) F-CHP staining for damaged collagen detec tion in the skin on POD7. Scale bars, 100 μm. (B) The three groups’ quantified F-CHP intensity (n = 6). (C) CD31 and α-SMA IF staining in the flap on POD7. Scale bars, 50 μm. (D) Quantified CD31/α-SMA-positive blood vessel density among the three groups (n = 6). (E) Dead cells in flap tissue sections on POD7 under the detection of TUNEL staining. Scale bar, 20 μm. (F) TUNEL-positive cell percentage in the dermal layer, quantified for each of the three groups (n = 6). (G) DHE staining of the skin tissues on POD7 among the three groups. Scale bars, 50 μm. (H) Quantified DHE among the three groups (n = 6). SEM error bars are used. Significance (*): p value < 0.05; equal variances ANOVA with LSD post hoc analysis or unequal variances Dunnett’s T3 technique

Article Snippet: The primary antibodies utilized were specific to CD31 (1:200), CD68 (1:200), α-SMA (1:200; Proteintech, 67735-1-Ig), iNOS (1:200), and Arg1 (1:200).

Techniques: Staining, TUNEL Assay

Journal: International Journal of Molecular Medicine

Article Title: Botulinum toxin type A prevents the phenotypic transformation of fibroblasts induced by TGF-β1 via the PTEN/PI3K/Akt signaling pathway

doi: 10.3892/ijmm.2019.4226

Figure Lengend Snippet: Effect of BTXA on the viability of mouse L929 fibroblasts. (A) The morphological changes of fibroblasts observed under a microscope. (B) The vimentin identification of fibroblasts with Hoechst 33258 was determined by immunofluorescence assay. (C) BTXA inhibited fibroblast viability, which was detected by cell counting kit-8 assay in a dose- (0, 0.125, 0.25, 0.5, 1 and 2 IU/ml) and time- (12, 24 and 48 h) dependent manner. ** P<0.01 vs. control. BTXA, botulinum toxin type A.

Article Snippet: After being blocked in bovine serum albumin (BSA), the fibroblasts were incubated first with anti-α-SMA antibody (A03744, 1:200; BosterBio) overnight at 4°C and then with Cy3-conjugated goat anti-rabbit secondary antibodies (BA1032, 1:500; Beyotime) for 0.5 h. Subsequently, the fibroblasts were stained with 4′,6-diamidino-2-phenylindole (DAPI; Beyotime) and images were captured under a fluorescence microscopy (Nikon).

Techniques: Microscopy, Immunofluorescence, Cell Counting, Control

Journal: International Journal of Molecular Medicine

Article Title: Botulinum toxin type A prevents the phenotypic transformation of fibroblasts induced by TGF-β1 via the PTEN/PI3K/Akt signaling pathway

doi: 10.3892/ijmm.2019.4226

Figure Lengend Snippet: BTXA suppresses fibroblast viability and promotes apoptosis. (A) BTXA suppressed the high fibroblast viability induced by TGF-β1. (B) BTXA increased the apoptosis of fibroblasts induced by 10 ng/ml of TGF-β1. Dotted line separation represents whether or not fibroblasts were treated with TGF-β1. Data are shown as the means ± SD, n=3. * P<0.05 and ** P<0.01 vs. control without TGF-β1; ^ P<0.05 and ^^ P<0.01 vs. control with TGF-β1. BTXA, botulinum toxin type A; TGF-β1, transforming growth factor-β1.

Article Snippet: After being blocked in bovine serum albumin (BSA), the fibroblasts were incubated first with anti-α-SMA antibody (A03744, 1:200; BosterBio) overnight at 4°C and then with Cy3-conjugated goat anti-rabbit secondary antibodies (BA1032, 1:500; Beyotime) for 0.5 h. Subsequently, the fibroblasts were stained with 4′,6-diamidino-2-phenylindole (DAPI; Beyotime) and images were captured under a fluorescence microscopy (Nikon).

Techniques: Control

Journal: International Journal of Molecular Medicine

Article Title: Botulinum toxin type A prevents the phenotypic transformation of fibroblasts induced by TGF-β1 via the PTEN/PI3K/Akt signaling pathway

doi: 10.3892/ijmm.2019.4226

Figure Lengend Snippet: BTXA prevents extracellular matrix (ECM) over-deposition in fibroblasts. BTXA decreased the high expression levels of collagen I, collagen III and α-SMA induced by 10 ng/ml of TGF-β1 in fibroblasts. The mRNA and protein levels of collagen I, collagen III and α-SMA were detected by (A) RT-qPCR and (B) western blot analysis, respectively. (C) α-SMA expression was determined by immunofluorescence. β-actin was used as an internal control. Dotted line separation represents whether or not fibroblast were treated with TGF-β1. Data were shown as the means ± SD, n=3. * P<0.05 and ** P<0.01 vs. control without TGF-β1; ^ P<0.05 and ^^ P<0.01 vs. control with TGF-β1. BTXA, botulinum toxin type A; TGF-β1, transforming growth factor-β1.

Article Snippet: After being blocked in bovine serum albumin (BSA), the fibroblasts were incubated first with anti-α-SMA antibody (A03744, 1:200; BosterBio) overnight at 4°C and then with Cy3-conjugated goat anti-rabbit secondary antibodies (BA1032, 1:500; Beyotime) for 0.5 h. Subsequently, the fibroblasts were stained with 4′,6-diamidino-2-phenylindole (DAPI; Beyotime) and images were captured under a fluorescence microscopy (Nikon).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Control

Journal: International Journal of Molecular Medicine

Article Title: Botulinum toxin type A prevents the phenotypic transformation of fibroblasts induced by TGF-β1 via the PTEN/PI3K/Akt signaling pathway

doi: 10.3892/ijmm.2019.4226

Figure Lengend Snippet: BTXA suppresses the expression levels of MMP-2 and MMP-9. BTXA decreased the high expression levels of MMP-2 and MMP-9 induced by 10 ng/ml of TGF-β1 in fibroblasts. The mRNA and protein levels of MMP-2 and MMP-9 were respectively detected by (A) RT-qPCR and (B) western blot analysis, respectively. β-actin was used as an internal control. Dotted line separation represents whether or not fibroblast were treated with TGF-β1. Data are shown as the means ± SD, n=3. * P<0.05 and ** P<0.01 vs. control without TGF-β1; ^ P<0.05 and ^^ P<0.01 vs. control with TGF-β1. BTXA, botulinum toxin type A; TGF-β1, transforming growth factor-β1; MMP, matrix metalloproteinase.

Article Snippet: After being blocked in bovine serum albumin (BSA), the fibroblasts were incubated first with anti-α-SMA antibody (A03744, 1:200; BosterBio) overnight at 4°C and then with Cy3-conjugated goat anti-rabbit secondary antibodies (BA1032, 1:500; Beyotime) for 0.5 h. Subsequently, the fibroblasts were stained with 4′,6-diamidino-2-phenylindole (DAPI; Beyotime) and images were captured under a fluorescence microscopy (Nikon).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control

Journal: International Journal of Molecular Medicine

Article Title: Botulinum toxin type A prevents the phenotypic transformation of fibroblasts induced by TGF-β1 via the PTEN/PI3K/Akt signaling pathway

doi: 10.3892/ijmm.2019.4226

Figure Lengend Snippet: Effect of BTXA on PTEN and caspase-3 expression, as well as PTEN methylation. (A) Effect of treatment of fibroblasts, which were cultured with or without 10 ng/ml of TGF-β1, with various concentrations (0.25, 0.5 and 1 UI/ml) of BTXA on the mRNA expression of PTEN, as determined by RT-qPCR. (B) Effects of BTXA on the protein levels of PTEN, pro-caspase-3 and cleaved-caspase-3, as detected by western blot analysis. (C) PTEN methylation was measured by methylation-specific PCR (MSP). M, methylated; U, unmethylated; MC, methylated control; UC, unmethylated control. β-actin was used as an internal control. Dotted line separation represents whether or not fibroblast were treated with TGF-β1. Data are shown as the means ± SD, n=3. * P<0.05 and ** P<0.01 vs. control without TGF-β1; ^ P<0.05 and ^^ P<0.01 vs. control with TGF-β1. BTXA, botulinum toxin type A; TGF-β1, transforming growth factor-β1; PTEN, phosphatase and tensin homolog deleted on chromosome ten.

Article Snippet: After being blocked in bovine serum albumin (BSA), the fibroblasts were incubated first with anti-α-SMA antibody (A03744, 1:200; BosterBio) overnight at 4°C and then with Cy3-conjugated goat anti-rabbit secondary antibodies (BA1032, 1:500; Beyotime) for 0.5 h. Subsequently, the fibroblasts were stained with 4′,6-diamidino-2-phenylindole (DAPI; Beyotime) and images were captured under a fluorescence microscopy (Nikon).

Techniques: Expressing, Methylation, Cell Culture, Quantitative RT-PCR, Western Blot, Control

Journal: International Journal of Molecular Medicine

Article Title: Botulinum toxin type A prevents the phenotypic transformation of fibroblasts induced by TGF-β1 via the PTEN/PI3K/Akt signaling pathway

doi: 10.3892/ijmm.2019.4226

Figure Lengend Snippet: BTXA inhibits the activities of DNMTs. (A) The high DNMT activity induced by 10 ng/ml of TGF-β1 was decreased by BTXA. (B and C) The mRNA and protein levels of DNMT1, DNMT3a and DNMT3b were determined by RT-qPCR and western blot analysis, respectively. β-actin was used as an internal control. Dotted line separation represents whether or not fibroblasts were treated with TGF-β1. Data are shown as the means ± SD, n=3. * P<0.05 and ** P<0.01 vs. control without TGF-β1; ^ P<0.05 and ^^ P<0.01 vs. control with TGF-β1. BTXA, botulinum toxin type A; TGF-β1, transforming growth factor-β1; DNMT, DNA methyltransferase.

Article Snippet: After being blocked in bovine serum albumin (BSA), the fibroblasts were incubated first with anti-α-SMA antibody (A03744, 1:200; BosterBio) overnight at 4°C and then with Cy3-conjugated goat anti-rabbit secondary antibodies (BA1032, 1:500; Beyotime) for 0.5 h. Subsequently, the fibroblasts were stained with 4′,6-diamidino-2-phenylindole (DAPI; Beyotime) and images were captured under a fluorescence microscopy (Nikon).

Techniques: Activity Assay, Quantitative RT-PCR, Western Blot, Control

Journal: International Journal of Molecular Medicine

Article Title: Botulinum toxin type A prevents the phenotypic transformation of fibroblasts induced by TGF-β1 via the PTEN/PI3K/Akt signaling pathway

doi: 10.3892/ijmm.2019.4226

Figure Lengend Snippet: BTXA inactivates the phosphorylation of PI3K and Akt. (A) The protein expression of p-PI3K, PI3K, p-Akt and Akt was examined by western blot analysis. (B) BTXA notably suppressed the ratio of p-PI3K/PI3K. (C) BTXA notably suppressed the ratio of p-Akt/Akt. β-actin was used as an internal control. Dotted line separation represents whether or not fibroblasts were treated with TGF-β1. Data are shown as the means ± SD, n=3. * P<0.05 and ** P<0.01 vs. control without TGF-β1; ^^ P<0.01 vs. control with TGF-β1.

Article Snippet: After being blocked in bovine serum albumin (BSA), the fibroblasts were incubated first with anti-α-SMA antibody (A03744, 1:200; BosterBio) overnight at 4°C and then with Cy3-conjugated goat anti-rabbit secondary antibodies (BA1032, 1:500; Beyotime) for 0.5 h. Subsequently, the fibroblasts were stained with 4′,6-diamidino-2-phenylindole (DAPI; Beyotime) and images were captured under a fluorescence microscopy (Nikon).

Techniques: Phospho-proteomics, Expressing, Western Blot, Control

Journal: Revista da Associacao Medica Brasileira (1992)

Article Title: Positive outcomes of phosphodiesterase type 5 inhibitor on histopathologic and biochemical changes induced by ureteral obstruction.

doi: 10.1590/1806-9282.65.3.388

Figure Lengend Snippet: FIGURE 2: A. Tissue analysis of α –smooth muscle (α-SMA) and B. Transforming growth factor-β (TGF-β) expressions in ureters using ELISA. The α-SMA and TGF-β expression levels were up-regulated in the PUUO and CUUO groups; however, they were down-regulated in the PUUO+T and CUUO+T groups ( * P < 0.0001 versus control).

Article Snippet: In the supernates, α-SMA and TGF-β levels were assayed using appropriate enzyme-linked immunosorbent assay (ELISA) kits (Cusabio CSB-E14027r and Boster EK0514, respectively).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Control