Journal: Biomaterials Research

Article Title: Fibroblast Activation Protein Alpha-Targeted Nanoparticles for Tumor Microenvironment Remodeling and Antitumor Therapy in Triple-Negative Breast Cancer

doi: 10.34133/bmr.0347

Figure Lengend Snippet: NP-FAP-DOX in vivo TME modulation. (A) Fap-α expression in tumors at the treatment endpoint was determined by RT-qPCR normalized to PBS ( n = 12 per group). Mean ± SEM. (B) Representative images of immunofluorescence staining of alpha-smooth muscle actin (α-SMA) in paraffin-embedded tumor sections at the treatment endpoint. Blue: nucleus, green: α-SMA. Scale bar: 50 μm. (C) Representative images of Masson’s trichrome staining in paraffin-embedded tumor sections at the treatment endpoint. Blue: collagen, pink: fibrin, red: muscle fibers, black: nucleus. Scale bar: 50 μm. (D) Representative second-harmonic generation (SHG) microscopy images of tumor sections showing fibrillar collagen structure. (E to H) Percentage of (E) T lymphocytes (CD45 + /CD3ε + ), (F) cytotoxic T cells (CD45 + /CD3ε + /CD8a + ), (G) M1-like macrophages (CD45 + /F4/80 + /CD86 + ), and (H) M2-like macrophages (CD45 + /F4/80 + /CD206 + ) in xenograft tumors analyzed by flow cytometry at the treatment endpoint ( n = 12 per group). (I) Ratios of M1 to M2 (M1/M2) were calculated as the percentage of M1-like macrophages divided by the percentage of M2-like macrophages on xenograft tumors analyzed by flow cytometry at the treatment endpoint ( n = 12 per group). Mean ± SEM. * P < 0.05, ** P < 0.01.

Article Snippet: Real-time quantitative polymerase chain reaction (RT-qPCR) was performed in a QuantStudio 5 Real-Time PCR System (Thermo Fisher Scientific, MA, USA) using 2 μl of resulting complementary DNA (cDNA), 5 μl of TaqMan Universal Master Mix (Applied Biosystems), 0.5 μl of TaqMan 20× assays (human FAP-α : Hs00990791_m1, human GAPDH : Hs03929097_g1, mouse Fap-α : Mm01329177_m1, mouse Gapdh : Mm99999915_g1, mouse Mki67 : Mm01278617_m1, mouse Myh7 : Mm00600555_m1; Applied Biosystems), and 2.5 μl of RNase-free water, according to the manufacturer’s instructions.

Techniques: In Vivo, Expressing, Quantitative RT-PCR, Immunofluorescence, Staining, Microscopy, Flow Cytometry

Journal: Biomaterials Research

Article Title: Fibroblast Activation Protein Alpha-Targeted Nanoparticles for Tumor Microenvironment Remodeling and Antitumor Therapy in Triple-Negative Breast Cancer

doi: 10.34133/bmr.0347

Figure Lengend Snippet: NP-FAP-DOX cardio and systemic toxicity evaluation. (A) Schematic representation of the in vivo study. 4T1 cells were injected into the mammary fat pad of BALB/C mice. The tumors, with an initial size of approximately 50 mm 3 , were intravenously treated with PBS (vehicle) or 50 mg/kg of nanoparticles (NPs: NP-CTR-DOX or NP-FAP-DOX) 4 times per week for 15 d ( n = 12 per group). Free doxorubicin (DOX) (3.75 mg/kg) was intravenously administered for 8 d. The mice were sacrificed 3 d after the last treatment, and the tumor and organs were collected. (B) Tumor growth curve mice treated with PBS (vehicle), NP-FAP-DOX, and free DOX ( n = 12 per group; mean ± SEM) (C) Body weight changes in tumor-bearing mice during treatment with PBS (vehicle), NP-FAP-DOX, or free DOX (mean ± SEM). (D) Myh7 expression in heart tissue at the treatment endpoint was determined by RT-qPCR and normalized to PBS (mean ± SEM). (E) Representative images of hematoxylin and eosin staining in paraffin-embedded heart, liver, spleen, and kidney tissue sections at the treatment endpoint. Thin arrows indicate cytoplasmic lysosomes. Thick arrows indicate degeneration of hepatocytes. Dashed areas in the spleen represent the white pulp. Dotted areas in the kidney represent the glomerular capsule, the asterisk represents tubule dilations, and the hash sign represents cellular infiltrations. Scale bar: 50 μm (heart, liver, and kidney) or 200 μm (spleen). DOX: Free doxorubicin. ****P < 0.0001.

Article Snippet: Real-time quantitative polymerase chain reaction (RT-qPCR) was performed in a QuantStudio 5 Real-Time PCR System (Thermo Fisher Scientific, MA, USA) using 2 μl of resulting complementary DNA (cDNA), 5 μl of TaqMan Universal Master Mix (Applied Biosystems), 0.5 μl of TaqMan 20× assays (human FAP-α : Hs00990791_m1, human GAPDH : Hs03929097_g1, mouse Fap-α : Mm01329177_m1, mouse Gapdh : Mm99999915_g1, mouse Mki67 : Mm01278617_m1, mouse Myh7 : Mm00600555_m1; Applied Biosystems), and 2.5 μl of RNase-free water, according to the manufacturer’s instructions.

Techniques: In Vivo, Injection, Expressing, Quantitative RT-PCR, Staining

Journal: Archives of biochemistry and biophysics

Article Title: Down-regulation of the mitochondrial fusion protein Opa1/Mfn2 promotes cardiomyocyte hypertrophy in Su5416/hypoxia-induced pulmonary hypertension rats.

doi: 10.1016/j.abb.2023.109743

Figure Lengend Snippet: Fig. 1. Bioinformatics analysis of MAMs-related gene expression in RV tissue of SuHx rats. A. The sample normalized diagram of the GSE42579 dataset. B. The principal component analysis (PCA) diagram of the GSE42579 dataset. C. Volcano map of differ entially expressed MAMs-related genes. D. The expression of Opa1, Mfn1, Mfn2, Dnm1l (Drp1), Nppa (Anp), Nppb (Bnp), and Myh7 in the GSE42579 dataset. **P < 0.01 vs Control. Opa1, optic atrophy 1; Mfn1, mitofusin 1; Mfn2, mitofusin 2; Dnm1l, dynamin-1-like protein, also named Drp1; Nppa, natriuretic peptides A, also named Anp; Nppb, natri uretic peptides B, also named Bnp; Myh7, myosin heavy chain 7.

Article Snippet: The PVDF membranes were incubated with primary antibodies against Opa1 (27733-1-AP, 1:1000, Proteintech, USA), Mfn2 (12186-1-AP, 1:1000, Proteintech, USA), Anp (AF7608, 1:1000, Beyotime, China), Bnp (AF6336, 1:1000, Beyotime, China), Myh7 (sc-53090, 1:100, Santa Cruz, USA) and α-tubulin (sc-5286, 1:500, Santa Cruz, USA) overnight at 4 ◦C.

Techniques: Gene Expression, Expressing, Control

Journal: Archives of biochemistry and biophysics

Article Title: Down-regulation of the mitochondrial fusion protein Opa1/Mfn2 promotes cardiomyocyte hypertrophy in Su5416/hypoxia-induced pulmonary hypertension rats.

doi: 10.1016/j.abb.2023.109743

Figure Lengend Snippet: Fig. 5. Overexpression of Opa1 or Mfn2 inhibited hypoxia-induced hypertrophy of H9c2 cells. A-B. The expression and distribution of Opa1 or Mfn2 in H9c2 cells were detected by immunofluorescence staining. C. Representative images of H9c2 cell morphology under the fluorescence microscope. D. Statistical graph of fluorescence intensity for Opa1. E. Statistical graph of fluorescence intensity for Mfn2. F. Statistical results of the cross-sectional area of H9c2 cells. G-L. Opa1, Mfn2, Anp, Bnp, and Myh7 protein expression in RV tissue. Data are expressed as mean ± SD. **P < 0.01 vs Control; ##P < 0.01, #P < 0.05 vs Hypoxia.

Article Snippet: The PVDF membranes were incubated with primary antibodies against Opa1 (27733-1-AP, 1:1000, Proteintech, USA), Mfn2 (12186-1-AP, 1:1000, Proteintech, USA), Anp (AF7608, 1:1000, Beyotime, China), Bnp (AF6336, 1:1000, Beyotime, China), Myh7 (sc-53090, 1:100, Santa Cruz, USA) and α-tubulin (sc-5286, 1:500, Santa Cruz, USA) overnight at 4 ◦C.

Techniques: Over Expression, Expressing, Immunofluorescence, Staining, Fluorescence, Microscopy, Control

Journal: Scientific reports

Article Title: An essential role for Wnt/β-catenin signaling in mediating hypertensive heart disease.

doi: 10.1038/s41598-018-27064-2

Figure Lengend Snippet: Figure 1. Chronic Ang II infusion induces cardiac hypertrophy and activates Wnt/β-catenin signaling. (A) Histological staining (H.E.) revealed overt cardiac hypertrophy in SD rats after 4 weeks of Ang II infusion. Scale bar, 50 µm. (B) Western blot analysis showed an increased expression of hypertrophic markers such as β-MHC and α-actin in the heart after Ang II infusion. Whole cardiac lysates were immunoblotted with antibodies against β-MHC, α-actin and α-tubulin, respectively. (C and D) Quantitative data on β-MHC and α-actin levels in different groups as indicated. *P < 0.05 versus controls (n = 4). (E) Quantitative, real-time RT-PCR (qRT- PCR) showed mRNA induction of multiple Wnt ligands in the heart of SD rat after 4 weeks of Ang II infusion. *P < 0.05 versus controls (n = 4). (F) Immunohistochemical staining for Wnt3a protein in the heart after Ang II infusion. Arrow indicates positive staining. Scale bar, 20 µm. (G) Representative micrographs showed induction of β-catenin protein in the heart after Ang II infusion. Yellow arrow indicates positive cardiomyocyte, whereas black arrow denotes positive cardiac fibroblast. Scale bar, 50 µm. (H–J) Western blot analysis showed cardiac β-catenin and active β-catenin protein expression in the heart after Ang II infusion. Representative Western blot (H) and quantitative data (I,J) are presented. (K–M) Western blot analysis showed the protein expression of β-catenin downstream target genes such as PAI-1 and Snail1. Representative Western blot (K) and quantitative data (L,M) are presented. *P < 0.05 versus controls (n = 4). Ctrl, controls. Ang II, angiotensin II.

Article Snippet: The primary antibodies used were as follows: β-MHC (MAB90961; R&D systems), α-actin (KM9006T; Sungene Biotech, Tianjin, China), β-catenin (610154; BD biosciences, San Jose, CA), active β-catenin (05-665; EMD Millipore, Billerica, MA), PAI-1 (AF1786; R&D systems, Minneapolis, MN), Snail1 (ab180714; Abcam, Cambridge, UK), fibronectin (F3648; Sigma, Darmstadt, Germany), α-SMA (clone 1A4; Sigma, Darmstadt, Germany), collagen I (BA0325; Boster, Pleasanton, CA), c-Myc (D84C12; CST, Massachusetts, USA) and α-tubulin (RM2007; Ray antibody Biotech, Beijing, China).

Techniques: Staining, Western Blot, Expressing, Quantitative RT-PCR, Immunohistochemical staining

Journal: Scientific reports

Article Title: An essential role for Wnt/β-catenin signaling in mediating hypertensive heart disease.

doi: 10.1038/s41598-018-27064-2

Figure Lengend Snippet: Figure 2. Inhibition of β-catenin signaling ameliorates Ang II-induced cardiac hypertrophy. (A) Representative micrographs showed immunohistochemical staining for cardiac β-MHC and α-actin proteins in various groups as indicated. Arrows indicate positive staining. Scale bar, 50 µm (top panel) and 20 µm (bottom panel). (B–D) Western blot analysis of β-MHC and α-actin in the heart after various treatments as indicated. Representative Western blots (B) and quantitative data for β-MHC (C) and α-actin (D) are presented. *P < 0.05 versus controls; †P < 0.05 versus Ang II alone (n = 5). (E) Ratio of heart weight/body weight of SD rats in various groups as indicated. HW, heart weight. BW, body weight. Ctrl, sham controls; Veh., vehicle; ICG, ICG-001; Los., losartan. *P < 0.05 versus sham controls; †P < 0.05 versus Ang II alone (n = 5).

Article Snippet: The primary antibodies used were as follows: β-MHC (MAB90961; R&D systems), α-actin (KM9006T; Sungene Biotech, Tianjin, China), β-catenin (610154; BD biosciences, San Jose, CA), active β-catenin (05-665; EMD Millipore, Billerica, MA), PAI-1 (AF1786; R&D systems, Minneapolis, MN), Snail1 (ab180714; Abcam, Cambridge, UK), fibronectin (F3648; Sigma, Darmstadt, Germany), α-SMA (clone 1A4; Sigma, Darmstadt, Germany), collagen I (BA0325; Boster, Pleasanton, CA), c-Myc (D84C12; CST, Massachusetts, USA) and α-tubulin (RM2007; Ray antibody Biotech, Beijing, China).

Techniques: Inhibition, Immunohistochemical staining, Staining, Western Blot

Journal: Scientific reports

Article Title: An essential role for Wnt/β-catenin signaling in mediating hypertensive heart disease.

doi: 10.1038/s41598-018-27064-2

Figure Lengend Snippet: Figure 7. Inhibition of β-catenin by ICG-001 prevents Ang II-induced cardiomyocyte hypertrophy in vitro. (A) Representative micrographs showed rhodamine staining of cardiomyocytes stimulated by Ang II in the absence or presence of ICG-001 or losartan. Scale bar, 50 µm. (B) Graphic presentation showed the relative sizes of cardiomyocytes in various groups as indicated. *P < 0.05 versus controls; †P < 0.05 versus Ang II alone (n = 15). (C) Representative Western blot analysis showed the protein levels of α-actin and β-MHC in cardiomyocytes after various treatments as indicated. (D,E) Graphic presentation showed the relative levels of α-actin and β-MHC proteins. *P < 0.05 versus controls; †P < 0.05 versus Ang II alone (n = 5). (F–H) qRT- PCR demonstrated the mRNA levels of ANP, BNP and β-MHC in cardiomyocytes after various treatments as indicated. (I–K) Western blots showed that recombinant Wnt3a induced α-actin and β-MHC expression in primary cardiomyocytes. Rat cardiomyocytes were treated with Wnt3a (100 ng/ml) for 24 hours. Western blot (I) and quantitative data (J and K) are presented. Ctrl, controls. Veh, Vehicle. ICG, ICG-001. Los, Losartan. *P < 0.05 versus controls; †P < 0.05 versus Ang II alone (n = 5).

Article Snippet: The primary antibodies used were as follows: β-MHC (MAB90961; R&D systems), α-actin (KM9006T; Sungene Biotech, Tianjin, China), β-catenin (610154; BD biosciences, San Jose, CA), active β-catenin (05-665; EMD Millipore, Billerica, MA), PAI-1 (AF1786; R&D systems, Minneapolis, MN), Snail1 (ab180714; Abcam, Cambridge, UK), fibronectin (F3648; Sigma, Darmstadt, Germany), α-SMA (clone 1A4; Sigma, Darmstadt, Germany), collagen I (BA0325; Boster, Pleasanton, CA), c-Myc (D84C12; CST, Massachusetts, USA) and α-tubulin (RM2007; Ray antibody Biotech, Beijing, China).

Techniques: Inhibition, In Vitro, Staining, Western Blot, Quantitative RT-PCR, Recombinant, Expressing

Journal: Data in Brief

Article Title: Supporting dataset of two integration-free induced pluripotent stem cell lines from related human donors

doi: 10.1016/j.dib.2021.107140

Figure Lengend Snippet: Reagents details.

Article Snippet: Differentiation Marker , Myosin Heavy Chain (MHC) REA VioBlue , 1:11 , Miltenyi 130-106-252; AB_2652979.

Techniques: Immunocytochemistry, Marker, Control

Journal: Pharmaceuticals (Basel, Switzerland)

Article Title: Oral Administration of Nacre Extract from Pearl Oyster Shells Has Anti-Aging Effects on Skin and Muscle, and Extends the Lifespan in SAMP8 Mice.

doi: 10.3390/ph17060713

Figure Lengend Snippet: Figure 6. Effect of nacre extract (NE) on the expression levels of forkhead box O (FOXO) 1, myosin heavy chain (MYH) 2, and MYH7 in the skeletal muscle. (a) The expression levels of FOXO1 in skeletal muscle were analyzed using real-time PCR. (b) Immunohistochemical staining of the skeletal muscle for FOXO1. Scale bar, 100 µm. (c) The stained areas of FOXO1 in skeletal muscle were measured. (d) Immunohistochemical staining of the skeletal muscle for MYH2 and MYH7. Scale bar, 100 µm. (e) The stained areas of MYH2 and MYH7 in skeletal muscle were measured. Ten to twenty different sections from five mice were analyzed for immunostaining and are presented as the mean ± SD. Data from five mice are presented as the mean ± SD for real-time PCR. Different letters indicate statistically significant differences (p < 0.05).

Article Snippet: Antibodies against sirtuin1, FOXO1, PGC1-α, myosin heavy chain (MYH) 2, MYH7, cyclindependent kinase inhibitor 2A (p16), cyclin-dependent kinase inhibitor 1 (p21), and succinate dehydrogenase (SDH) were purchased from Biorbyt (San Francisco, CA, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Immunohistochemical staining, Staining, Immunostaining

Journal: Regenerative Therapy

Article Title: Assessment of human bioengineered cardiac tissue function in hypoxic and re-oxygenized environments to understand functional recovery in heart failure

doi: 10.1016/j.reth.2021.03.007

Figure Lengend Snippet: PCR primer information.

Article Snippet: MYH7 , Hs01110632_m1.

Techniques: TaqMan Assay