Journal: Journal of Nanobiotechnology

Article Title: Design of RGD-functionalized GSH-responsive pegylated polymeric protacs for selective BRD4 degradation and EndMT-driven cardiac fibrosis inhibition

doi: 10.1186/s12951-026-04036-7

Figure Lengend Snippet: RGD-PEG-MZ1 exerted a profound inhibitory effect on the EndMT in vitro. ( A-D ) The expression of Collagen-I, Fibronectin, VE-cadherin, α-SMA and FSP1 was assessed using western blotting (WB) in HUVECs and HMEC-1. ( E ) Immunofluorescence staining for α-SMA (green) and VE-cadherin (green) in HUVECs. Scale bar, 100 μm. The data are presented as the Mean ± SD of three independent experiments. # p < 0.05 and ## p < 0.01 vs. NC group; ns, p > 0.05 vs. control group; * p < 0.05 and ** p < 0.01 vs. control group (one-way ANOVA followed by Bonferroni post hoc test)

Article Snippet: The slides were initially stabilized with a 5% goat serum solution, then exposed to primary antibodies—α-SMA (Proteintech, 14395-1-AP), Collagen-I (Proteintech, 14695-1-AP), CD31 (Proteintech, 66065-2-Ig), Raf1 (Abcam, ab181115), Raf1 (phospho S259) (Abcam, ab173539), ERK1 + ERK2 (Abcam, ab184699) and ERK1 (phospho T202 + Y204) + ERK2 (phospho T185 + Y187) (Abcam, ab278538)—and subsequently incubated at 4 °C for an extended period.

Techniques: In Vitro, Expressing, Western Blot, Immunofluorescence, Staining, Control

Journal: iScience

Article Title: The role of long noncoding RNA Nron in atherosclerosis development and plaque stability

doi: 10.1016/j.isci.2022.103978

Figure Lengend Snippet:

Article Snippet: Primary antibodies specific for F4/80 (ab100790, Abcam), α-SMA (ab240654, Abcam) or VEGFA (ab1316, Abcam) was used.

Techniques: Virus, Recombinant, Plasmid Preparation, Protease Inhibitor, Bicinchoninic Acid Protein Assay, Silver Staining, In Vitro, TUNEL Assay, Software

Journal: EMBO Molecular Medicine

Article Title: Characterization and therapy of fertilization failure in murine and human models with HNRNPR mutations

doi: 10.1038/s44321-026-00374-z

Figure Lengend Snippet: ( A ) MII oocyte and zygote fluorescence imaging reflecting fertilization after ICSI of Hnrnpr -mutated sperm. Formation of male and female pronuclei indicates successful fertilization. Tubulin labels the spindle, showing that the oocyte is at metaphase of meiosis II, and P1 marks the first polar body. The white dotted line defines the boundary of the zygotes. Scale bar = 10 μm. ( B ) Quantification of the percentage of zygotes from ( A ). Data are presented as mean ± s.d., and P values were calculated using a two-sided Student’s t test. Data in ( A , B ) represent results from six independent experiments (two technical and three biological replicates, n = 6).

Article Snippet: Rabbit anti-alpha Tubulin , MedChemExpress , Cat# HY- P86200.

Techniques: Fluorescence, Imaging

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