Journal: Inflammation

Article Title: Caveolin-2 Knockdown Alleviated Sepsis-induced Acute Lung Injury Via Promoting Macrophage M2 Polarization and Inhibiting Apoptosis by Hippo Signaling Pathway

doi: 10.1007/s10753-025-02378-2

Figure Lengend Snippet: Cav2 knockdown inhibited pulmonary apoptosis and promoted M2 polarization of sepsis-induced ALI in vivo. ( A ) Western blotting analysis of the expression of Bax, Bak, Caspase-3, Bcl-2, CD86, iNOS, CD80, CD206, CD163 and Arg1 protein in lung tissues, GAPDH serves as an internal control ( N = 3 per group). ( B , C , D , E ) The mRNA expression of Bax, Bak, Caspase-3 and Bcl-2 protein in lung tissues, detected by RT-qPCR ( N = 8 per group). ( F, G , H , I , J , K ) RT-qPCR analysis of the expression of CD86, iNOS, CD80, CD206, Arg1 and CD163 in lung tissues ( N = 8 per group). Mice were euthanized 24 h post-CLP, above data of animal models were expressed as median and interquartile range. * P < 0.05, ** P < 0.01, *** P < 0.001vs. Control group. # P < 0.05, ## P < 0.01 vs. CLP group

Article Snippet: Antibodies against Bax (14796S, CST, USA, dilution ratio of WB,1:1000, IHC,1:200), Bak (12150S, CST, USA, dilution ratio of WB,1:1000, IHC, 1:200), Caspase-3 (9662S, CST, USA, dilution ratio of WB,1:1000, IHC, 1:1000), Bcl-2 (15071S, CST, USA, dilution ratio of WB,1:1000, IHC, 1:500), Lats1 (ab243656, Abcam, USA, dilution ratio of WB,1:1000), Mst1 (ab317410, Abcam, UK,dilution ratio of WB,1:1000), Sav1 (ab307698, Abcam, UK,dilution ratio of WB,1:1000), p -YAP (ab313464, Abcam, USA, dilution ratio of WB,1:500), YAP (HY- P86386 , MCE, USA, dilution ratio of WB,1:1000, IF,1:200), p -TAZ (Sc-17610-R, Santa Cruz, USA, dilution ratio of WB,1:200), TAZ (83669S, CST, USA, dilution ratio of WB,1:1000, IHC, 1:200), iNOS (22226-1-AP, Proteintech, China, dilution ratio of WB,1:1000), CD86 (13395-1-AP, Proteintech, China,dilution ratio of WB,1:2000), CD80 (66406-1-lg, Proteintech, China, dilution ratio of WB,1:1000), CD163 (HY- P81177 , MCE, USA, dilution ratio of WB,1:2000), CD206 (32647-1-AP, Proteintech, China, dilution ratio of WB,1:2000), Arg1 (16001-1-AP, Proteintech, China, dilution ratio of WB,1:5000), GAPDH (2118S, CST, USA, dilution ratio of WB,1:1000), Histone H3 (HY- P86672 , MCE, USA, dilution ratio of WB,1:10000), VT02956 (MCE, USA), MY-875 (MCE, USA), HRP-conjugated AffiniPure Goat Anti-Rabbit IgG H&L (HY-P8001, MCE, USA, dilution ratio of WB,1:8000), HRP-conjugated AffiniPure Goat Anti-Mouse IgG H&L (HY-P8004, MCE, USA,dilution ratio of WB,1:10000), APC-anti CD80 antibody (17-0801-82, Thermo Fisher Scientific, USA, dilution ratio of Flow,1:100), APC-anti CD206 (17-2061-82, Thermo Fisher Scientific, USA, dilution ratio of Flow,1:100), PE-MHC II (12-5322-81, Thermo Fisher Scientific, USA, dilution ratio of Flow,1:200), FITC-anti-CD163 (11-1631-82, Thermo Fisher Scientific, USA, dilution ratio of Flow,1:50).

Techniques: Knockdown, In Vivo, Western Blot, Expressing, Control, Quantitative RT-PCR

Journal: Inflammation

Article Title: Caveolin-2 Knockdown Alleviated Sepsis-induced Acute Lung Injury Via Promoting Macrophage M2 Polarization and Inhibiting Apoptosis by Hippo Signaling Pathway

doi: 10.1007/s10753-025-02378-2

Figure Lengend Snippet: Cav2 knockdown mitigated macrophage apoptosis and promoted M2 polarization in MHS cells induced by LPS. ( A ) Western blotting analysis the expression of Bak, Bax, Caspase-3, Bcl-2, iNOS, CD86, CD80, CD206, Arg1 and CD163 proteins in MHS cells, GAPDH serves as an internal control ( N = 3 per group). ( B , C , D , E ) RT-qPCR analysis the mRNA expression of Bak, Bax, Caspase-3 and Bcl-2 protein in MHS cells ( N = 8 per group). ( F , G , H , I , J , K ) RT-qPCR analysis of the mRNA expression of CD86, iNOS, CD80, CD206, Arg1 and CD163 in MHS cells ( N = 8 per group). ( L , M ) Flow cytometry was used to detect M1 and M2 polarization related phenotype in each MHS cells groups ( N = 3 per group). Data were expressed as mean ± SD, * P < 0.05, ** P < 0.01 vs. Control group. # P < 0.05, ## P < 0.01, ### P < 0.001vs. LPS group

Article Snippet: Antibodies against Bax (14796S, CST, USA, dilution ratio of WB,1:1000, IHC,1:200), Bak (12150S, CST, USA, dilution ratio of WB,1:1000, IHC, 1:200), Caspase-3 (9662S, CST, USA, dilution ratio of WB,1:1000, IHC, 1:1000), Bcl-2 (15071S, CST, USA, dilution ratio of WB,1:1000, IHC, 1:500), Lats1 (ab243656, Abcam, USA, dilution ratio of WB,1:1000), Mst1 (ab317410, Abcam, UK,dilution ratio of WB,1:1000), Sav1 (ab307698, Abcam, UK,dilution ratio of WB,1:1000), p -YAP (ab313464, Abcam, USA, dilution ratio of WB,1:500), YAP (HY- P86386 , MCE, USA, dilution ratio of WB,1:1000, IF,1:200), p -TAZ (Sc-17610-R, Santa Cruz, USA, dilution ratio of WB,1:200), TAZ (83669S, CST, USA, dilution ratio of WB,1:1000, IHC, 1:200), iNOS (22226-1-AP, Proteintech, China, dilution ratio of WB,1:1000), CD86 (13395-1-AP, Proteintech, China,dilution ratio of WB,1:2000), CD80 (66406-1-lg, Proteintech, China, dilution ratio of WB,1:1000), CD163 (HY- P81177 , MCE, USA, dilution ratio of WB,1:2000), CD206 (32647-1-AP, Proteintech, China, dilution ratio of WB,1:2000), Arg1 (16001-1-AP, Proteintech, China, dilution ratio of WB,1:5000), GAPDH (2118S, CST, USA, dilution ratio of WB,1:1000), Histone H3 (HY- P86672 , MCE, USA, dilution ratio of WB,1:10000), VT02956 (MCE, USA), MY-875 (MCE, USA), HRP-conjugated AffiniPure Goat Anti-Rabbit IgG H&L (HY-P8001, MCE, USA, dilution ratio of WB,1:8000), HRP-conjugated AffiniPure Goat Anti-Mouse IgG H&L (HY-P8004, MCE, USA,dilution ratio of WB,1:10000), APC-anti CD80 antibody (17-0801-82, Thermo Fisher Scientific, USA, dilution ratio of Flow,1:100), APC-anti CD206 (17-2061-82, Thermo Fisher Scientific, USA, dilution ratio of Flow,1:100), PE-MHC II (12-5322-81, Thermo Fisher Scientific, USA, dilution ratio of Flow,1:200), FITC-anti-CD163 (11-1631-82, Thermo Fisher Scientific, USA, dilution ratio of Flow,1:50).

Techniques: Knockdown, Western Blot, Expressing, Control, Quantitative RT-PCR, Flow Cytometry

Journal: Inflammation

Article Title: Caveolin-2 Knockdown Alleviated Sepsis-induced Acute Lung Injury Via Promoting Macrophage M2 Polarization and Inhibiting Apoptosis by Hippo Signaling Pathway

doi: 10.1007/s10753-025-02378-2

Figure Lengend Snippet: Cav2 knockdown promoted M2 polarization and attenuated macrophage apoptosis by regulating Hippo signaling pathway in vitro. ( A ) Immunofluorescence analysis assessed the nuclear expression and localization of Nucleus YAP and Nucleus TAZ. ( B ) Western blotting analysis the expression of Nucleus YAP, Nucleus TAZ, Lats1, Mst1, Sav1, Cytoplasm p-YAP, Cytoplasm p-TAZ, Bak, Bax, Caspase-3, Bcl-2, iNOS, CD86, CD80, CD206, Arg1 and CD163 proteins in MHS cells, Histone H3 and GAPDH serves as an internal control of nuclear and cytoplasmic proteins ( N = 3 per group). ( B , C ) The expression of M1 and M2 polarization related phenotype in MHS cells, detected by flow cytometry. ( D , E , F , G ) The mRNA expression of Bak, Bax, Caspase-3 and Bcl-2. ( H , I , J , K ) The mRNA expression of iNOS, CD86, CD206 and Arg1. ( L , M , N ) The mRNA expression of IL-1β, IL-6 and TNF-α in MHS cells, detected by RT-qPCR. Data were expressed as mean ± SD, * P < 0.05, ** P < 0.01 vs. Control group. # P < 0.05, ## P < 0.01, ### P < 0.001 vs. LPS group. $ P < 0.05 vs. LPS + LV-si-Cav2 group. Ɛ P < 0.05, ƐƐ P < 0.01 vs. LPS + LV-OE-Cav2 group

Article Snippet: Antibodies against Bax (14796S, CST, USA, dilution ratio of WB,1:1000, IHC,1:200), Bak (12150S, CST, USA, dilution ratio of WB,1:1000, IHC, 1:200), Caspase-3 (9662S, CST, USA, dilution ratio of WB,1:1000, IHC, 1:1000), Bcl-2 (15071S, CST, USA, dilution ratio of WB,1:1000, IHC, 1:500), Lats1 (ab243656, Abcam, USA, dilution ratio of WB,1:1000), Mst1 (ab317410, Abcam, UK,dilution ratio of WB,1:1000), Sav1 (ab307698, Abcam, UK,dilution ratio of WB,1:1000), p -YAP (ab313464, Abcam, USA, dilution ratio of WB,1:500), YAP (HY- P86386 , MCE, USA, dilution ratio of WB,1:1000, IF,1:200), p -TAZ (Sc-17610-R, Santa Cruz, USA, dilution ratio of WB,1:200), TAZ (83669S, CST, USA, dilution ratio of WB,1:1000, IHC, 1:200), iNOS (22226-1-AP, Proteintech, China, dilution ratio of WB,1:1000), CD86 (13395-1-AP, Proteintech, China,dilution ratio of WB,1:2000), CD80 (66406-1-lg, Proteintech, China, dilution ratio of WB,1:1000), CD163 (HY- P81177 , MCE, USA, dilution ratio of WB,1:2000), CD206 (32647-1-AP, Proteintech, China, dilution ratio of WB,1:2000), Arg1 (16001-1-AP, Proteintech, China, dilution ratio of WB,1:5000), GAPDH (2118S, CST, USA, dilution ratio of WB,1:1000), Histone H3 (HY- P86672 , MCE, USA, dilution ratio of WB,1:10000), VT02956 (MCE, USA), MY-875 (MCE, USA), HRP-conjugated AffiniPure Goat Anti-Rabbit IgG H&L (HY-P8001, MCE, USA, dilution ratio of WB,1:8000), HRP-conjugated AffiniPure Goat Anti-Mouse IgG H&L (HY-P8004, MCE, USA,dilution ratio of WB,1:10000), APC-anti CD80 antibody (17-0801-82, Thermo Fisher Scientific, USA, dilution ratio of Flow,1:100), APC-anti CD206 (17-2061-82, Thermo Fisher Scientific, USA, dilution ratio of Flow,1:100), PE-MHC II (12-5322-81, Thermo Fisher Scientific, USA, dilution ratio of Flow,1:200), FITC-anti-CD163 (11-1631-82, Thermo Fisher Scientific, USA, dilution ratio of Flow,1:50).

Techniques: Knockdown, In Vitro, Immunofluorescence, Expressing, Western Blot, Control, Flow Cytometry, Quantitative RT-PCR

Journal: Inflammation

Article Title: Caveolin-2 Knockdown Alleviated Sepsis-induced Acute Lung Injury Via Promoting Macrophage M2 Polarization and Inhibiting Apoptosis by Hippo Signaling Pathway

doi: 10.1007/s10753-025-02378-2

Figure Lengend Snippet: Cav2 knockdown promoted M2 polarization and attenuated macrophage apoptosis by regulating Hippo signaling pathway in vivo. ( A ) Western blotting analysis the expression of Nucleus TAZ, Nucleus YAP, Lats1, Mst1, Sav1, Cytoplasm p-YAP Cytoplasm p-TAZ, Bak, Bax, Caspase-3, Bcl-2, iNOS, CD86, CD80, CD206,Arg1 and CD163 proteins in lung tissues, Histone H3 and GAPDH serves as an internal control of nuclear and cytoplasmic proteins ( N = 3 per group). ( B ) IHC results showing the expression levels of Bak, Bax, Caspase-3 and Bcl-2 (200X, Scale bar = 50 μm). ( C , D ) Percentage of IHC positive cells and positive cells counts score of IHC ( N = 8 per group). ( E ) HE staining of lung tissues (100X, Scale bar = 100 μm). ( F ) Lung tissues of HE staining injury scoring ( N = 8 per group). ( G , H , I ) The mRNA expression of IL-1β, IL-6 and TNF-α in lung tissues, detected by RT-qPCR ( N = 8 per group). Above data of animal models were expressed as median and interquartile range, * P < 0.05, ** P < 0.01, *** P < 0.001vs.Control group. # P < 0.05, ## P < 0.0 vs.CLP group. $ P < 0.05, $$ P < 0.01 vs. CLP + AAV-si-Cav2 group

Article Snippet: Antibodies against Bax (14796S, CST, USA, dilution ratio of WB,1:1000, IHC,1:200), Bak (12150S, CST, USA, dilution ratio of WB,1:1000, IHC, 1:200), Caspase-3 (9662S, CST, USA, dilution ratio of WB,1:1000, IHC, 1:1000), Bcl-2 (15071S, CST, USA, dilution ratio of WB,1:1000, IHC, 1:500), Lats1 (ab243656, Abcam, USA, dilution ratio of WB,1:1000), Mst1 (ab317410, Abcam, UK,dilution ratio of WB,1:1000), Sav1 (ab307698, Abcam, UK,dilution ratio of WB,1:1000), p -YAP (ab313464, Abcam, USA, dilution ratio of WB,1:500), YAP (HY- P86386 , MCE, USA, dilution ratio of WB,1:1000, IF,1:200), p -TAZ (Sc-17610-R, Santa Cruz, USA, dilution ratio of WB,1:200), TAZ (83669S, CST, USA, dilution ratio of WB,1:1000, IHC, 1:200), iNOS (22226-1-AP, Proteintech, China, dilution ratio of WB,1:1000), CD86 (13395-1-AP, Proteintech, China,dilution ratio of WB,1:2000), CD80 (66406-1-lg, Proteintech, China, dilution ratio of WB,1:1000), CD163 (HY- P81177 , MCE, USA, dilution ratio of WB,1:2000), CD206 (32647-1-AP, Proteintech, China, dilution ratio of WB,1:2000), Arg1 (16001-1-AP, Proteintech, China, dilution ratio of WB,1:5000), GAPDH (2118S, CST, USA, dilution ratio of WB,1:1000), Histone H3 (HY- P86672 , MCE, USA, dilution ratio of WB,1:10000), VT02956 (MCE, USA), MY-875 (MCE, USA), HRP-conjugated AffiniPure Goat Anti-Rabbit IgG H&L (HY-P8001, MCE, USA, dilution ratio of WB,1:8000), HRP-conjugated AffiniPure Goat Anti-Mouse IgG H&L (HY-P8004, MCE, USA,dilution ratio of WB,1:10000), APC-anti CD80 antibody (17-0801-82, Thermo Fisher Scientific, USA, dilution ratio of Flow,1:100), APC-anti CD206 (17-2061-82, Thermo Fisher Scientific, USA, dilution ratio of Flow,1:100), PE-MHC II (12-5322-81, Thermo Fisher Scientific, USA, dilution ratio of Flow,1:200), FITC-anti-CD163 (11-1631-82, Thermo Fisher Scientific, USA, dilution ratio of Flow,1:50).

Techniques: Knockdown, In Vivo, Western Blot, Expressing, Control, Staining, Quantitative RT-PCR

Journal: Clinical and Experimental Pharmacology & Physiology

Article Title: Mechanisms of IL‐17A Neutralisation in Alleviating Renal Fibrosis and Inflammation in Spontaneously Hypertensive Rats

doi: 10.1111/1440-1681.70116

Figure Lengend Snippet: Effects of IL‐17A neutralisation on macrophage polarisation in SHR renal tissues. (A) Representative IHC staining of M1 macrophage markers (iNOS and CD86) with quantitative analysis of their positive areas. (B) Representative IHC staining of M2 macrophage markers (Arg‐1 and CD163) with quantitative analysis of their positive areas. (C) Representative immunoblots and relative expression levels of iNOS, CD86, Arg‐1, and CD163 proteins. (D) Proportion of CD86 + and CD163 + cells among CD68 + macrophages. (E) mRNA expression levels of iNOS, CD86, Arg‐1, and CD163. Data are presented as mean ± SD ( n = 6).

Article Snippet: Sections were then incubated overnight at 4°C with primary antibodies against: E‐cadherin (Boster, China), Collagen III (Boster, China), inducible nitric oxide synthase (iNOS) (Boster, China), CD86 (Boster, China), arginase‐1 (Arg‐1) (Boster, China), CD163 (Boster, China), α‐smooth muscle actin (α‐SMA) (Cell Signalling Technology, USA).

Techniques: Immunohistochemistry, Western Blot, Expressing

Journal: Acta biomaterialia

Article Title: A subtype specific probe for targeted magnetic resonance imaging of M2 tumor-associated macrophages in brain tumors.

doi: 10.1016/j.actbio.2025.01.003

Figure Lengend Snippet: Fig. 6. Immunohistochemical and FACS validations of M2pep-uIONP targeting M2 TAMs. Confocal microscopic images of tumor tissues collected from mice at 24 h after i.v. injection of probes show (A) Cy7-M2pep-uIONP were highly co-localizing with CD68- and CD163-double positive M2 TAM, revealing the targeting specificity. In comparison, the non-targeted control probe Cy7-scM2pep-uIONP and Cy7-Ferumoxytol not only exhibited much less tumoral accumulation, but also were not co-localized with M2 TAM. Scale bars: 37 µm. (B) Flow cytometric zebra plots and (C) the corresponding quantification of CD68- and CD163-double positive M2 TAM targeted by FITC-M2pep-uIONP, FITC-scM2pep-uIONP, and FITC-Ferumoxytol. The gating strategy involves sorting CD163+ M2 TAM population in live cells dissociated from mouse cerebrums by gating on CD68+ pan-macrophage populations, followed by further gating to assess the FITC-labeled nanoparticles.

Article Snippet: Rabbit polyclonal anti-CD163 antibody was purchased from Boster Biological Technology (Pleasanton, CA, USA).

Techniques: Immunohistochemical staining, Injection, Comparison, Control, Labeling

Journal: Experimental and Therapeutic Medicine

Article Title: Role of SARS‑CoV‑2 nucleocapsid protein in affecting immune cells and insights on its molecular mechanisms

doi: 10.3892/etm.2023.12203

Figure Lengend Snippet: Quantification of TNF-α, IL-10, IL-6 and CD163. The concentrations of (A) TNF-α, (B) IL-10, (C) IL-6 and (D) CD163 were determined in the supernatant of cultured Raw264.7 cells treated with the severe acute respiratory syndrome coronavirus 2 N protein. Data are presented as mean ± SD (n=3). * P<0.05 and ** P<0.01 vs. blank control.

Article Snippet: PageRuler TM Prestained protein ladder (cat. no. 26616) was purchased from Thermo Fisher Scientific, Inc. Trypsin-EDTA (0.25%) used to digest Raw264.7 cells for cell passage was purchased from Gibco; Thermo Fisher Scientific Inc. TNF-α Elisa kit (Cat. no. EK0527), IL-6 Elisa Kit (Cat. no. EK0411), IL-10 Elisa Kit (Cat. no. EK0417) and CD163 ELISA kit (Cat. no. EK1146) were purchased from Boster Biological Technology co., Ltd.).

Techniques: Cell Culture, Control

Journal: Human molecular genetics

Article Title: Tweak regulates astrogliosis, microgliosis and skeletal muscle atrophy in a mouse model of amyotrophic lateral sclerosis.

doi: 10.1093/hmg/ddv094

Figure Lengend Snippet: Figure 2. Tweak induces death of primary motoneurons via CD163, endocytosis and caspase-3. (A) Motoneurons were cultured for 24 h and treated with indicated

Article Snippet: The ELISA kits used were as follows: mouse IL-6 (R&D Systems), mouse Tweak and mouse CD163 (CUSABIO Biotech Co).

Techniques: Cell Culture