cd163 proteintech  (Proteintech)

Journal: Scientific reports

Article Title: ANGPTL4 induces Kupffer cell M2 polarization to mitigate acute rejection in liver transplantation.

doi: 10.1038/s41598-024-81832-x

Figure Lengend Snippet: Fig. 2. ANGPTL4 expression in the liver following transplantation. (A) RT-qPCR evaluation of ANGPTL4 levels in rat liver tissues. (B) ELISA results of serum ANGPTL4 protein levels. (C) Immunohistochemical staining of ANGPTL4 in liver graft sections (200x magnification). ns P > 0.05, *P < 0.05; **P < 0.01; ***P < 0.001; n = 6.

Article Snippet: Comparisons between two groups were made using an unpaired Student’s t-test, Name Dilution Supplier Code GAPDH WB: 1:10000 Proteintech 60004-1-Ig Arg1 WB: 1:1000 Proteintech 16001-1-AP CD86 WB: 1:1000, IHC: 1:1000 Zenbio 380,350 P65 WB: 1:1000 Proteintech 66535-1-Ig p-P65 WB: 1:1000, IF: 1:500 CST 3033 IKKβ WB: 1:1000 ABclonal A2087 p-IKKβ WB: 1:1000 Proteintech A2087 IκBα WB: 1:1000 ABclonal A19714 p-IκBα WB: 1:1000 ABclonal AP0707 Goat anti-rabbit WB: 1:20000 Zenbio 550,018 CD68 IF: 1:100 Invitrogen MA5-13324 CD163 IHC: 1:1000 Proteintech 16646-1-AP ANGPTL4 IHC: 1:1000 Proteintech 18374-1-AP Table 2.

Techniques: Expressing, Transplantation Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining

Journal: Scientific reports

Article Title: ANGPTL4 induces Kupffer cell M2 polarization to mitigate acute rejection in liver transplantation.

doi: 10.1038/s41598-024-81832-x

Figure Lengend Snippet: Fig. 3. ANGPTL4 recombinant protein mitigates AR following liver transplantation. (A, B) Representative HE images of liver grafts taken 7 days post-transplantation at original magnifications of ×100 and ×400, along with RAI scoring of the liver. (C) TUNEL staining to identify apoptotic cells in liver tissues across different groups post-transplantation (200× magnification). (D) Measurement of serum liver enzyme levels after transplantation using a microtiter assay. (E) ELISA evaluation of serum inflammatory factors in rats following liver transplantation. ns P > 0.05, *P < 0.05; **P < 0.01; ***P < 0.001; n = 6.

Article Snippet: Comparisons between two groups were made using an unpaired Student’s t-test, Name Dilution Supplier Code GAPDH WB: 1:10000 Proteintech 60004-1-Ig Arg1 WB: 1:1000 Proteintech 16001-1-AP CD86 WB: 1:1000, IHC: 1:1000 Zenbio 380,350 P65 WB: 1:1000 Proteintech 66535-1-Ig p-P65 WB: 1:1000, IF: 1:500 CST 3033 IKKβ WB: 1:1000 ABclonal A2087 p-IKKβ WB: 1:1000 Proteintech A2087 IκBα WB: 1:1000 ABclonal A19714 p-IκBα WB: 1:1000 ABclonal AP0707 Goat anti-rabbit WB: 1:20000 Zenbio 550,018 CD68 IF: 1:100 Invitrogen MA5-13324 CD163 IHC: 1:1000 Proteintech 16646-1-AP ANGPTL4 IHC: 1:1000 Proteintech 18374-1-AP Table 2.

Techniques: Recombinant, Transplantation Assay, TUNEL Assay, Staining, Enzyme-linked Immunosorbent Assay

Journal: Scientific reports

Article Title: ANGPTL4 induces Kupffer cell M2 polarization to mitigate acute rejection in liver transplantation.

doi: 10.1038/s41598-024-81832-x

Figure Lengend Snippet: Fig. 4. ANGPTL4 facilitates the in vivo shift of KCs towards the M2 phenotype. (A) WB analysis of polarization-related protein expression in KCs, with quantitative analysis based on band intensity. (B) Immunohistochemical staining of CD86 and CD163. ns P > 0.05, *P < 0.05; **P < 0.01; ***P < 0.001; n = 6.

Article Snippet: Comparisons between two groups were made using an unpaired Student’s t-test, Name Dilution Supplier Code GAPDH WB: 1:10000 Proteintech 60004-1-Ig Arg1 WB: 1:1000 Proteintech 16001-1-AP CD86 WB: 1:1000, IHC: 1:1000 Zenbio 380,350 P65 WB: 1:1000 Proteintech 66535-1-Ig p-P65 WB: 1:1000, IF: 1:500 CST 3033 IKKβ WB: 1:1000 ABclonal A2087 p-IKKβ WB: 1:1000 Proteintech A2087 IκBα WB: 1:1000 ABclonal A19714 p-IκBα WB: 1:1000 ABclonal AP0707 Goat anti-rabbit WB: 1:20000 Zenbio 550,018 CD68 IF: 1:100 Invitrogen MA5-13324 CD163 IHC: 1:1000 Proteintech 16646-1-AP ANGPTL4 IHC: 1:1000 Proteintech 18374-1-AP Table 2.

Techniques: In Vivo, Expressing, Immunohistochemical staining, Staining

Journal: Scientific reports

Article Title: ANGPTL4 induces Kupffer cell M2 polarization to mitigate acute rejection in liver transplantation.

doi: 10.1038/s41598-024-81832-x

Figure Lengend Snippet: Fig. 5. ANGPTL4 recombinant protein promotes M2 polarization of KCs. (A) RT-qPCR and ELISA were used to measure IL-10, IL-1β, and TNF-α expression levels in KCs isolated from rat liver and their corresponding levels in the cell supernatant. (B) WB analysis of polarization-related protein levels in KCs, with quantitative evaluation based on band intensity. (C) Flow cytometry was employed to assess M1-polarized KCs, using FITC-labeled CD68 for KC identification and APC-labeled CD86 to detect M1 polarization. ns P > 0.05, *P < 0.05; **P < 0.01; ***P < 0.001; n = 3.

Article Snippet: Comparisons between two groups were made using an unpaired Student’s t-test, Name Dilution Supplier Code GAPDH WB: 1:10000 Proteintech 60004-1-Ig Arg1 WB: 1:1000 Proteintech 16001-1-AP CD86 WB: 1:1000, IHC: 1:1000 Zenbio 380,350 P65 WB: 1:1000 Proteintech 66535-1-Ig p-P65 WB: 1:1000, IF: 1:500 CST 3033 IKKβ WB: 1:1000 ABclonal A2087 p-IKKβ WB: 1:1000 Proteintech A2087 IκBα WB: 1:1000 ABclonal A19714 p-IκBα WB: 1:1000 ABclonal AP0707 Goat anti-rabbit WB: 1:20000 Zenbio 550,018 CD68 IF: 1:100 Invitrogen MA5-13324 CD163 IHC: 1:1000 Proteintech 16646-1-AP ANGPTL4 IHC: 1:1000 Proteintech 18374-1-AP Table 2.

Techniques: Recombinant, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Expressing, Isolation, Flow Cytometry, Labeling

Journal: Scientific reports

Article Title: ANGPTL4 induces Kupffer cell M2 polarization to mitigate acute rejection in liver transplantation.

doi: 10.1038/s41598-024-81832-x

Figure Lengend Snippet: Fig. 6. Hepatocyte-derived ANGPTL4 is crucial for suppressing inflammatory activation in KCs. (A) Schematic of the experimental procedure: co-culture of HCs with LPS-stimulated KCs. (B) ELISA analysis of ANGPTL4 levels in the KCs culture medium. (C) RT-qPCR and ELISA analysis of IL-10, IL-1β, and TNF-α expression levels. (D) WB analysis of polarization-related proteins in KCs, with quantitative assessment based on band intensity. (E) Flow cytometry analysis of M1-polarized cells, identifying KCs with FITC-labeled CD68 and M1 markers with APC-labeled CD86. (F) Western blot analysis of proteins related to the NF-κB pathway, with quantification based on band intensity. (G) Immunofluorescence staining to localize phosphorylated p65. ns P > 0.05, *P < 0.05; **P < 0.01; ***P < 0.001; n = 3.

Article Snippet: Comparisons between two groups were made using an unpaired Student’s t-test, Name Dilution Supplier Code GAPDH WB: 1:10000 Proteintech 60004-1-Ig Arg1 WB: 1:1000 Proteintech 16001-1-AP CD86 WB: 1:1000, IHC: 1:1000 Zenbio 380,350 P65 WB: 1:1000 Proteintech 66535-1-Ig p-P65 WB: 1:1000, IF: 1:500 CST 3033 IKKβ WB: 1:1000 ABclonal A2087 p-IKKβ WB: 1:1000 Proteintech A2087 IκBα WB: 1:1000 ABclonal A19714 p-IκBα WB: 1:1000 ABclonal AP0707 Goat anti-rabbit WB: 1:20000 Zenbio 550,018 CD68 IF: 1:100 Invitrogen MA5-13324 CD163 IHC: 1:1000 Proteintech 16646-1-AP ANGPTL4 IHC: 1:1000 Proteintech 18374-1-AP Table 2.

Techniques: Derivative Assay, Activation Assay, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Expressing, Flow Cytometry, Labeling, Western Blot, Immunofluorescence, Staining

Journal: Scientific reports

Article Title: ANGPTL4 induces Kupffer cell M2 polarization to mitigate acute rejection in liver transplantation.

doi: 10.1038/s41598-024-81832-x

Figure Lengend Snippet: Fig. 7. Knockdown of ANGPTL4 expression in HCs within the co-culture system promotes M1 polarization of KCs. (A) Schematic representation of the experimental procedure: siRNA-mediated knockdown of ANGPTL4 in HCs co-cultured with LPS-stimulated KCs. (B) RT-qPCR and ELISA assessments of IL-10, IL-1β, and TNF-α levels. (C) WB analysis of polarization-related proteins in KCs, with quantification of band intensity. (D) Flow cytometry of M1-type polarized cells, with FITC-labeled CD68 and APC-labeled CD86. (E) Western blot assessment of proteins associated with the NF-κB pathway, including band intensity quantification. (F) Immunofluorescence staining to identify the localization of phosphorylated p65. ns P > 0.05, *P < 0.05; **P < 0.01; ***P < 0.001; n = 3.

Article Snippet: Comparisons between two groups were made using an unpaired Student’s t-test, Name Dilution Supplier Code GAPDH WB: 1:10000 Proteintech 60004-1-Ig Arg1 WB: 1:1000 Proteintech 16001-1-AP CD86 WB: 1:1000, IHC: 1:1000 Zenbio 380,350 P65 WB: 1:1000 Proteintech 66535-1-Ig p-P65 WB: 1:1000, IF: 1:500 CST 3033 IKKβ WB: 1:1000 ABclonal A2087 p-IKKβ WB: 1:1000 Proteintech A2087 IκBα WB: 1:1000 ABclonal A19714 p-IκBα WB: 1:1000 ABclonal AP0707 Goat anti-rabbit WB: 1:20000 Zenbio 550,018 CD68 IF: 1:100 Invitrogen MA5-13324 CD163 IHC: 1:1000 Proteintech 16646-1-AP ANGPTL4 IHC: 1:1000 Proteintech 18374-1-AP Table 2.

Techniques: Knockdown, Expressing, Co-Culture Assay, Cell Culture, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Labeling, Western Blot, Immunofluorescence, Staining

Journal: Parasite immunology

Article Title: SEA Alleviates Hepatic Ischaemia-Reperfusion Injury by Promoting M2 Macrophage Polarisation.

doi: 10.1111/pim.13061

Figure Lengend Snippet: FIGURE 4 | SEA pretreatment promoted the polarisation of M2 macrophages. Mice were subjected to 60 min of hepatic ischaemia, followed by 6 h of reperfusion. (A) Representatives of immunofluorescence staining of CD68 (n = 5 per group, five biological replicates from five individual animals; original magnification 100× and 200×, scale bar: 100 μm). (B) Representatives of immunofluorescence staining of CD163 (n = 5 per group, five biological replicates from five individual animals; original magnification 100× and 200×, scale bar: 100 μm). (C, D) The expression of M2 macrophage marker (CD206 and Arg1) in the liver was detected via qRT-PCR (n = 5 per group, five biological replicates from five individual animals). These results were obtained from at least three independent experiments. Values are presented as mean ± SEM. **p < 0.01, *p < 0.05, ns > 0.05.

Article Snippet: Then the sections were incubated with primary antibodies dissolved in 5% donkey serum solution containing 0.3% Triton at 4°C overnight (anti- MPO, Cat# ab208670, Abcam, 1:100; anti- CD68, Cat# ab53444, Abcam, 1:100; anti- CD163, Cat# 16646- 1- AP, Proteintech, 1:100).

Techniques: Immunofluorescence, Staining, Expressing, Marker, Quantitative RT-PCR