Journal: Molecular Medicine Reports

Article Title: Osthole inhibits proliferation of kainic acid-activated BV-2 cells by modulating the Notch signaling pathway

doi: 10.3892/mmr.2020.11455

Figure Lengend Snippet: Effect of osthole on TNF-α, IL-6 and iNOS contents in cultured BV-2 cells activated by kainic acid. The levels of (A) IL-6 (B) NOS2/iNOS and (C) TNF-α were detected using ELISA kits. Data are presented as the mean ± standard deviation of three independent experiments. *P<0.05 vs. the Con group, # P<0.05 vs. the KA group. IL, interleukin; NOS2, nitric oxide synthase 2; iNOS, inducible nitric oxide synthase; TNF-α, tumor necrosis factor-α; KA, kainic acid; Con, control.

Article Snippet: Rabbit monoclonal anti-mouse Notch-2 antibody (cat. no. 5732) was obtained from Cell Signaling Technology, Inc. Rabbit monoclonal anti-mouse Jagged-1 antibody (cat. no. E-AB-30148), mouse interleukin (IL)-6 ELISA kit (cat. no. E-EL-M0044c), mouse TNF-α ELISA kit (cat. no. E-EL-M0049c) and mouse nitric oxide synthase 2 (NOS2)/inducible i) NOS ELISA Kit (cat. no. E-EL-M0696c) were purchased from Elabscience.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Standard Deviation, Control

Journal: Experimental and Therapeutic Medicine

Article Title: Effects of sepsis on the metabolism of sphingomyelin and cholesterol in mice with liver dysfunction

doi: 10.3892/etm.2017.5226

Figure Lengend Snippet: IL-1β, TNF-α, ALT and AST levels in the serum of mice. (A) Plasma levels of IL-1β and TNF-α were analyzed using ELISA kits. (B) Serum ALT and AST levels were measured. Data are presented as the mean ± standard deviation (n=6). **P<0.001 vs. control group; #P<0.05 vs. LPS group. IL, interleukin; TNF, tumor necrosis factor; ALT, alanine transaminase; AST, aspartate transaminase; L + P, LPS + pyrrolidine dithiocarbamate; LPS, lipopolysaccharide.

Article Snippet: To identify the successful establishment of the sepsis model and liver dysfunction, plasma levels of interleukin (IL)-1β (E-EL-M0037c) and tumor necrosis factor (TNF)-α (E-EL-M0049c) (both from Elabscience Biotechnology Co., Ltd., Wuhan, China) were analyzed by ELISA using commercial kits, and the levels of alanine transaminase (ALT, C009-2) and aspartate transaminase (AST, C010-2) (both from Nanjing Jiancheng Bioengineering Institute, Nanjing, China) were detected using commercial kits.

Techniques: Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Fractalkine aggravates LPS‐induced macrophage activation and acute kidney injury via Wnt/β‐catenin signalling pathway

doi: 10.1111/jcmm.16707

Figure Lengend Snippet: FKN regulated polarization in LPS‐induced J774A.1 cells via Wnt/β‐catenin signalling. A, B, Western blot analysis and their respective quantitation were performed to detect the expression levels of iNOS, TNF‐α, ARG‐1 and IL‐10 protein in J774A.1 cells. C, D, ELISA was used to detect the levels of TNF‐α and ARG‐1 in the cell supernatants. * P < .05 compared with the control group; # P < .05 compared with the LPS group. E, F, Immunofluorescence analysis was used to ascertained the subcellular localization of iNOS and ARG‐1. Scale bars represent 10 μm

Article Snippet: Cyclin D1(SBJ‐M0517, Senbeijia Bioengineering Institute,), TNF‐α (E‐EL‐M0049c, Cusabio Biotech Co., Ltd) and ARG‐1 (CSB‐EL002005MO, Cusabio Biotech Co., Ltd) were detected according to manufacturer's instructions using ELISA Kits.

Techniques: Western Blot, Quantitation Assay, Expressing, Enzyme-linked Immunosorbent Assay, Control, Immunofluorescence

Journal: Journal of Cellular and Molecular Medicine

Article Title: Fractalkine aggravates LPS‐induced macrophage activation and acute kidney injury via Wnt/β‐catenin signalling pathway

doi: 10.1111/jcmm.16707

Figure Lengend Snippet: FKN deficiency suppressed macrophages polarization and proliferation in mice kidney tissues via inhibition of Wnt/β‐catenin signalling. A, B, Western blot analysis and their respective quantitation showing the protein expression of FKN, β‐catenin, Wnt‐4, c‐myc, cyclinD1, iNOS, TNF‐α, ARG‐1 and IL‐10 in mice kidney tissues. * P < .05 compared with the WT mice; # P < .05 compared with the LPS mice

Article Snippet: Cyclin D1(SBJ‐M0517, Senbeijia Bioengineering Institute,), TNF‐α (E‐EL‐M0049c, Cusabio Biotech Co., Ltd) and ARG‐1 (CSB‐EL002005MO, Cusabio Biotech Co., Ltd) were detected according to manufacturer's instructions using ELISA Kits.

Techniques: Inhibition, Western Blot, Quantitation Assay, Expressing

Journal: Animals : an Open Access Journal from MDPI

Article Title: Effect of Actin Alpha Cardiac Muscle 1 on the Proliferation and Differentiation of Bovine Myoblasts and Preadipocytes

doi: 10.3390/ani11123468

Figure Lengend Snippet: RT-qPCR primer sequence.

Article Snippet: The protein samples in the gel were transferred to a polyvinylidene fluoride membrane (PVDF, Merck Millipore, Taufkirchen, Germany), and then sealed with QuickBlock Western blocking solution (Beyotime Biotechnology, Shanghai, China) for 20 min. GAPDH (rabbit anti-GAPDH, 1:10,000 Abcam, Cambridge, UK, NP_001029206.1), ACTC1 (rabbit anti-ACTC1, 1:1000, Invitrogen, CA, USA NP_001029757.1), MYOD1 (mouse anti-MYOD1, 1:1000, Abcam, Cambridge, NT, UK, NP_001035568.2), MYOG (rabbit anti-MYOG, 1:1000, Abcam, Cambridge, NT, UK, NP_001104795.1), MYHC (mouse anti-MYHC, 1:200, Abcam, Cambridge, NT, UK), MRF4 (mouse anti-MYF6, 1:100, Santa Cruz Biotechnology, Santa Cruz, CA, USA, NP_861527.1), MEF2A (rabbit anti-MEF2A, 1:1000, Abcam, Cambridge, NT, UK, NP_001077107.1), FABP4 (rabbit anti-FABP4, 1:1000, Abcam, NT, UK, NP_776739.1), PPARγ (rabbit anti-PPARγ, 1:1000, Boster, Wuhan, China, NP_851367.1), FASN (rabbit anti-FASN, 1:2000, Abcam, Cambridge, NT, UK, NP_777087.1) antibodies were added and incubated overnight at 4 °C.

Techniques: Sequencing

Journal: Animals : an Open Access Journal from MDPI

Article Title: Effect of Actin Alpha Cardiac Muscle 1 on the Proliferation and Differentiation of Bovine Myoblasts and Preadipocytes

doi: 10.3390/ani11123468

Figure Lengend Snippet: Detection of adipogenic marker gene expression at mRNA level after ectopic expression of ACTC1 . ( a – e ) Detection of mRNA expression of ACTC1 , PPARγ , FABP4 , FASN and SCD1 genes on D0, D2, D4, D6 and D8 days by qRT-PCR after ectopic expression ACTC1 . Each experiment was performed in triplicate. Error bars represent s.e.m. * p < 0.05; ** p < 0.01.

Article Snippet: The protein samples in the gel were transferred to a polyvinylidene fluoride membrane (PVDF, Merck Millipore, Taufkirchen, Germany), and then sealed with QuickBlock Western blocking solution (Beyotime Biotechnology, Shanghai, China) for 20 min. GAPDH (rabbit anti-GAPDH, 1:10,000 Abcam, Cambridge, UK, NP_001029206.1), ACTC1 (rabbit anti-ACTC1, 1:1000, Invitrogen, CA, USA NP_001029757.1), MYOD1 (mouse anti-MYOD1, 1:1000, Abcam, Cambridge, NT, UK, NP_001035568.2), MYOG (rabbit anti-MYOG, 1:1000, Abcam, Cambridge, NT, UK, NP_001104795.1), MYHC (mouse anti-MYHC, 1:200, Abcam, Cambridge, NT, UK), MRF4 (mouse anti-MYF6, 1:100, Santa Cruz Biotechnology, Santa Cruz, CA, USA, NP_861527.1), MEF2A (rabbit anti-MEF2A, 1:1000, Abcam, Cambridge, NT, UK, NP_001077107.1), FABP4 (rabbit anti-FABP4, 1:1000, Abcam, NT, UK, NP_776739.1), PPARγ (rabbit anti-PPARγ, 1:1000, Boster, Wuhan, China, NP_851367.1), FASN (rabbit anti-FASN, 1:2000, Abcam, Cambridge, NT, UK, NP_777087.1) antibodies were added and incubated overnight at 4 °C.

Techniques: Marker, Gene Expression, Expressing, Quantitative RT-PCR

Journal: Animals : an Open Access Journal from MDPI

Article Title: Effect of Actin Alpha Cardiac Muscle 1 on the Proliferation and Differentiation of Bovine Myoblasts and Preadipocytes

doi: 10.3390/ani11123468

Figure Lengend Snippet: Detection of adipogenic marker gene expression at protein level after ectopic expression of ACTC1 . ( a ) Detect the proteins expression of ACTC1, PPARγ, FABP4, FASN and SCD1 on D2, D4, D6 and D8 days by Western blot after ectopic expression of ACTC1 . ( b – f ) Use Image J software to measure the gray value of protein bands. Each experiment was performed in triplicate. Error bars represent s.e.m. * p < 0.05; ** p < 0.01. Original western blot figures in .

Article Snippet: The protein samples in the gel were transferred to a polyvinylidene fluoride membrane (PVDF, Merck Millipore, Taufkirchen, Germany), and then sealed with QuickBlock Western blocking solution (Beyotime Biotechnology, Shanghai, China) for 20 min. GAPDH (rabbit anti-GAPDH, 1:10,000 Abcam, Cambridge, UK, NP_001029206.1), ACTC1 (rabbit anti-ACTC1, 1:1000, Invitrogen, CA, USA NP_001029757.1), MYOD1 (mouse anti-MYOD1, 1:1000, Abcam, Cambridge, NT, UK, NP_001035568.2), MYOG (rabbit anti-MYOG, 1:1000, Abcam, Cambridge, NT, UK, NP_001104795.1), MYHC (mouse anti-MYHC, 1:200, Abcam, Cambridge, NT, UK), MRF4 (mouse anti-MYF6, 1:100, Santa Cruz Biotechnology, Santa Cruz, CA, USA, NP_861527.1), MEF2A (rabbit anti-MEF2A, 1:1000, Abcam, Cambridge, NT, UK, NP_001077107.1), FABP4 (rabbit anti-FABP4, 1:1000, Abcam, NT, UK, NP_776739.1), PPARγ (rabbit anti-PPARγ, 1:1000, Boster, Wuhan, China, NP_851367.1), FASN (rabbit anti-FASN, 1:2000, Abcam, Cambridge, NT, UK, NP_777087.1) antibodies were added and incubated overnight at 4 °C.

Techniques: Marker, Gene Expression, Expressing, Western Blot, Software

Journal: Cell Death & Disease

Article Title: Pentraxin 3 regulated by miR-224-5p modulates macrophage reprogramming and exacerbates osteoarthritis associated synovitis by targeting CD32

doi: 10.1038/s41419-022-04962-y

Figure Lengend Snippet: A , B IF staining and quantification of F4/80 (upper), iNOS (middle), and CD206 (lower) in synovial tissues of DMM mice treated with vehicle, rmPTX3 or PTX3-NAb for 8 weeks, n = 5 per group. Scale bar: 50 µm. C Relative mRNA expression level of iNOS and ARG in LPS-, IL-4- or rmPTX3-treated RAW264.7 cells, n = 9 per group. D Relative mRNA expression level of IL-1β, IL-6, and TNF-α in rmPTX3-treated RAW264.7 cells, n = 9 per group. E ELISA of IL-1β, IL-6, and TNF-α in the supernatants of RAW264.7 treated with rmPTX3, n = 8 per group. * P < 0.05, ** P < 0.01, *** P < 0.001, ns not significant. Data are shown as means ± SD. Statistical analyses were conducted by one-way analysis of variance followed by Dunnett’s multiple comparison test ( B ), two-way analysis of variance followed by Sidak’s multiple comparison test ( C ) or unpaired t -test ( D and E ). Boxed area is enlarged in the top right corner.

Article Snippet: We used the Mouse IL-1β, IL-6 and TNF-α ELISA Kits (Elabscience Biotechnology, Bethesda, MD, USA, E-EL-M0037c, E-EL-M0044c and E-EL-M0049c) to analyze IL-1β, IL-6 and TNF-α levels in the supernatants of macrophage treated with 400 ng/mL rmPTX3.

Techniques: Staining, Expressing, Enzyme-linked Immunosorbent Assay, Comparison