Journal: Frontiers in Immunology

Article Title: Dynamic changes in human THP-1-derived M1-to-M2 macrophage polarization during Thelazia callipaeda MIF induction

doi: 10.3389/fimmu.2022.1078880

Figure Lengend Snippet: Polarization-related signaling pathway of macrophages was activated after T.cp -MIF treatment. The macrophages binded with TLR4, and T.cp -MIF activated NF-κB signaling pathway 24 hours after induction. At this time, macrophages showed M1 type polarization, while PI3K/AKT/mTOR signaling pathway is activated at 48 hours, making macrophages show M2 type polarization.

Article Snippet: After blocking, the corresponding primary antibodies such as TLR4 (Wuhan Sanying Biotechnology Co., Ltd.), NF-κB (Wuhan Sanying Biotechnology Co., Ltd.), p-NF-κB (Cell signaling technology, USA), PI3K (Wuhan Sanying Biotechnology Co., Ltd.), p-PI3K (Abcam, UK), Akt (Wuhan Sanying Biotechnology Co. Ltd.), and β-actin (Hangzhou Huaan Biotechnology Co., Ltd.) was separately added to the box (antibody concentrations were diluted according to the instructions), incubated at 4°C for over 12 h. After incubation, the membrane was washed 3 times with TBST buffer for 10 min each time.

Techniques:

Journal: Frontiers in Immunology

Article Title: Dynamic changes in human THP-1-derived M1-to-M2 macrophage polarization during Thelazia callipaeda MIF induction

doi: 10.3389/fimmu.2022.1078880

Figure Lengend Snippet: Effect of T.cp -MIF on the expression of macrophage polarization-related proteins in different time. (A–D) The protein expression levers of TLR4, NF-κBp65, PI3K and AKT were analyzed by western blot and normalized against β-actin expression. * indicated P <0.05. Results are showed as the mean of three times of independent experiments ( x̄ ±SD, n=3).

Article Snippet: After blocking, the corresponding primary antibodies such as TLR4 (Wuhan Sanying Biotechnology Co., Ltd.), NF-κB (Wuhan Sanying Biotechnology Co., Ltd.), p-NF-κB (Cell signaling technology, USA), PI3K (Wuhan Sanying Biotechnology Co., Ltd.), p-PI3K (Abcam, UK), Akt (Wuhan Sanying Biotechnology Co. Ltd.), and β-actin (Hangzhou Huaan Biotechnology Co., Ltd.) was separately added to the box (antibody concentrations were diluted according to the instructions), incubated at 4°C for over 12 h. After incubation, the membrane was washed 3 times with TBST buffer for 10 min each time.

Techniques: Expressing, Western Blot

Journal: Frontiers in Immunology

Article Title: Dynamic changes in human THP-1-derived M1-to-M2 macrophage polarization during Thelazia callipaeda MIF induction

doi: 10.3389/fimmu.2022.1078880

Figure Lengend Snippet: The effect of TAK-242 on the expression of macrophage polarization-related proteins. (A–D) The cells were pretreated with TAK-242 (the TLR4 inhibitor) for 6 h, and the expression levels of TLR4, NF-κBp65, p-NF-κBp65, PI3K, p-PI3K, Akt and p-Akt were detected by western blot. * indicated P <0.05. Results are showed as the mean of three times of independent experiments ( x̄ ±SD, n=3).

Article Snippet: After blocking, the corresponding primary antibodies such as TLR4 (Wuhan Sanying Biotechnology Co., Ltd.), NF-κB (Wuhan Sanying Biotechnology Co., Ltd.), p-NF-κB (Cell signaling technology, USA), PI3K (Wuhan Sanying Biotechnology Co., Ltd.), p-PI3K (Abcam, UK), Akt (Wuhan Sanying Biotechnology Co. Ltd.), and β-actin (Hangzhou Huaan Biotechnology Co., Ltd.) was separately added to the box (antibody concentrations were diluted according to the instructions), incubated at 4°C for over 12 h. After incubation, the membrane was washed 3 times with TBST buffer for 10 min each time.

Techniques: Expressing, Western Blot

Journal: Frontiers in Endocrinology

Article Title: Plasma-derived exosomal miRNAs as potentially novel biomarkers for type 2 diabetes mellitus with abdominal obesity

doi: 10.3389/fendo.2025.1656132

Figure Lengend Snippet: Exosome identification. The exosomes were analyzed by (A) TEM and NTA assay (B) . (C, D) The protein expression of CD9, CD81, TSG101, Calnexin, and FABP4 in exosomes was detected by Western blot assay. Cell lysate (CL) was used as a positive control for calmodulin.

Article Snippet: Western blot analysis was performed to determine the expression of CD9 (Wuhan Sanying, China, 1:2000)、CD81 (Wuhan Sanying, China, 1:1000)、Tsg101 (Wuhan Sanying, China, 1:2000)、Calnexin (Wuhan Sanying, China, 1:5000)、FABP4 (Santa Cruz, America, 1:500)、β-actin (Wuhan Sanying, China, 1:4000).

Techniques: Expressing, Western Blot, Positive Control

Journal: Discover Oncology

Article Title: The prognostic value of miR-323b-5p in non-small cell lung cancer and its mechanism of targeting IL1A in regulating the proliferation and cell cycle of non-small cell lung cancer cells

doi: 10.1007/s12672-026-04409-2

Figure Lengend Snippet: miR-323b-5p targets IL1A. A The intersection of predicted targets from TargetScan and miRDB databases. B The expression of IL1A and MEOX2 in A549, HCC827, NCI-H1299 and BEAS-2B cells. C The expression of IL1A in the serum of NSCLC patients ( n = 120) and healthy subjects ( n = 60). D The binding site of miR-323b-5p and IL1A. E Dual-luciferase reporter assay indicates that miR-323b-5p directly regulates IL1A. F Expression of IL1A in A549, HCC827, and NCI-H1299 cells after transfection with miR-323b-5p mimics. The experiment was repeated 3 times independently. ( p < 0.05*, p < 0.001***)

Article Snippet: The antibodies used include: IL1A (Wuhan Sanying, 1:1000), P21 (Wuhan Sanying, 1:1500), CCND1 (Wuhan Sanying, 1:5000), cyclin dependent kinase 4 (CDK4, Wuhan Sanying, 1: 1000), β-actin (Wuhan Sanying, 1:2000).

Techniques: Expressing, Binding Assay, Luciferase, Reporter Assay, Transfection

Journal: Discover Oncology

Article Title: The prognostic value of miR-323b-5p in non-small cell lung cancer and its mechanism of targeting IL1A in regulating the proliferation and cell cycle of non-small cell lung cancer cells

doi: 10.1007/s12672-026-04409-2

Figure Lengend Snippet: miR-323b-5p affects A549 cell proliferation and blocks the cell cycle through IL1A. After transfection or non-transfection of A549 cells with the control mimic (mimic NC), miR-323b-5p mimic (mimic MIR), IL1A overexpression plasmid (ovIL1A), control inhibitor (inhibitor NC), miR-323b-5p inhibitor (inhibitor MIR) and IL1A knockdown plasmid (siIL1A). The transfection efficiency of each group of cells was detected by RT-qPCR ( A ), cell proliferation was detected by CCK-8 ( B ), the cell cycle status of A549 cells was detected by flow cytometry ( C ), and the expression of P21, CCND1 and CDK4 mRNA ( D ) and protein ( E ) in the cells was detected by RT-qPCR and WB respectively. The experiment was repeated 3 times independently with 3 biological replicates ( n = 3). (*compared with Control, p < 0.05*. # compared with mimic NC, p < 0.05 # .)

Article Snippet: The antibodies used include: IL1A (Wuhan Sanying, 1:1000), P21 (Wuhan Sanying, 1:1500), CCND1 (Wuhan Sanying, 1:5000), cyclin dependent kinase 4 (CDK4, Wuhan Sanying, 1: 1000), β-actin (Wuhan Sanying, 1:2000).

Techniques: Transfection, Control, Over Expression, Plasmid Preparation, Knockdown, Quantitative RT-PCR, CCK-8 Assay, Flow Cytometry, Expressing

Journal: Discover Oncology

Article Title: The prognostic value of miR-323b-5p in non-small cell lung cancer and its mechanism of targeting IL1A in regulating the proliferation and cell cycle of non-small cell lung cancer cells

doi: 10.1007/s12672-026-04409-2

Figure Lengend Snippet: miR-323b-5p affects HCC827 cell proliferation and blocks the cell cycle through IL1A. After transfection or non-transfection of A549 cells with the control mimic (mimic NC), miR-323b-5p mimic (mimic MIR), IL1A overexpression plasmid (ovIL1A), control inhibitor (inhibitor NC), miR-323b-5p inhibitor (inhibitor MIR) and IL1A knockdown plasmid (siIL1A). The transfection efficiency of each group of cells was detected by RT-qPCR ( A ), cell proliferation was detected by CCK-8 ( B ), the cell cycle status of A549 cells was detected by flow cytometry ( C ), and the expression of P21, CCND1 and CDK4 mRNA ( D ) and protein ( E ) in the cells was detected by RT-qPCR and WB respectively. The experiment was repeated 3 times independently with 3 biological replicates ( n = 3). (*compared with Control, p < 0.001***. # compared with mimic NC, p < 0.001 ### . & compared with inhibitor NC, p < 0.05 & )

Article Snippet: The antibodies used include: IL1A (Wuhan Sanying, 1:1000), P21 (Wuhan Sanying, 1:1500), CCND1 (Wuhan Sanying, 1:5000), cyclin dependent kinase 4 (CDK4, Wuhan Sanying, 1: 1000), β-actin (Wuhan Sanying, 1:2000).

Techniques: Transfection, Control, Over Expression, Plasmid Preparation, Knockdown, Quantitative RT-PCR, CCK-8 Assay, Flow Cytometry, Expressing

Journal: Discover Oncology

Article Title: The prognostic value of miR-323b-5p in non-small cell lung cancer and its mechanism of targeting IL1A in regulating the proliferation and cell cycle of non-small cell lung cancer cells

doi: 10.1007/s12672-026-04409-2

Figure Lengend Snippet: miR-323b-5p affects NCI-H1299 cell proliferation and blocks the cell cycle through IL1A. After transfection or non-transfection of A549 cells with the control mimic (mimic NC), miR-323b-5p mimic (mimic MIR), IL1A overexpression plasmid (ovIL1A), control inhibitor (inhibitor NC), miR-323b-5p inhibitor (inhibitor MIR) and IL1A knockdown plasmid (siIL1A). The transfection efficiency of each group of cells was detected by RT-qPCR ( A ), cell proliferation was detected by CCK-8 ( B ), the cell cycle status of A549 cells was detected by flow cytometry ( C ), and the expression of P21, CCND1 and CDK4 mRNA ( D ) and protein ( E ) in the cells was detected by RT-qPCR and WB respectively. The experiment was repeated 3 times independently with 3 biological replicates ( n = 3). ( # compared with mimic NC, p < 0.001 ### .)

Article Snippet: The antibodies used include: IL1A (Wuhan Sanying, 1:1000), P21 (Wuhan Sanying, 1:1500), CCND1 (Wuhan Sanying, 1:5000), cyclin dependent kinase 4 (CDK4, Wuhan Sanying, 1: 1000), β-actin (Wuhan Sanying, 1:2000).

Techniques: Transfection, Control, Over Expression, Plasmid Preparation, Knockdown, Quantitative RT-PCR, CCK-8 Assay, Flow Cytometry, Expressing

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Uncovering the mechanisms of homologous point acupuncture on knee osteoarthritis through an integrated study of metabolomics and proteomics

doi: 10.3389/fbioe.2026.1791109

Figure Lengend Snippet: Effects of HAP on cartilage autophagy and apoptosis in KOA model rats. Note: (A,C–F) Expression of LC3 I, LC3 II, Beclin1, C-Caspase3 and Bax in cartilage tissue (GAPDH as internal control). (B,G,H) Expression of p62 and Bcl2 in cartilage (GAPDH as internal control). *** p < 0.001, ** p < 0.01, * p < 0.05.

Article Snippet: All the primary antibodies were validated by either the manufacturer or other available publications, listed as follows: HIF1α (Boster, A00013-1), HK2 (Abclonal, A0994), c-caspase3 (Affinity, AF7022), Bax (Wuhan Sanying, 50599-2-Ig), LC3 (CST, 12741T), Beclin1 (Wuhan Sanying, 11306-1-AP), and GAPDH (Wuhan Sanying, 60004-1-Ig) from WUHAN HUAYAN Biotechnology CO., LTD. NLRP3 (Affinity, BF8029), ASC (Affinity, DF6304), caspase1 (Affinity, AF5418), p62 (Wuhan Sanying, 18420-1-AP), Bcl2 (Wuhan Sanying, 60178-1-Ig) from WUHAN Fabre Biotechnology CO., LTD. After washing 5 times for 5 min each with TBST (T-Pro), and a horseradish peroxidase (HRP)-conjugated secondary antibody (Boster, BA1051 and BA1054) was applied for 2 h at room temperature.

Techniques: Expressing, Control