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: American Journal of Physiology - Cell Physiology

Article Title: Hemoglobin scavenger receptor CD163 as a potential biomarker of hemolysis-induced hepatobiliary injury in sickle cell disease

doi: 10.1152/ajpcell.00386.2023

Figure Lengend Snippet: Hemoglobin scavenger receptor CD163 is associated with hepatobiliary injury in sickle cell disease (SCD). A : representative images of immunofluorescence (IF) analysis of SCD patient biopsied liver samples show abundant CD163-positive cells. B , left : representative IF images (merged and single channels) of colocalization assay show higher percentage of CD163 colocalized with positive F4/80 staining of Kupffer cells in SCD mouse liver. Right : bar graph depicts the Pearson’s coefficient of colocalization in control (CON) and SCD mouse liver. C , left : representative IF images (merged and single channels) of colocalization assay show higher percentage of CD163 colocalized with positive CD11b staining of monocytes in SCD mouse liver. Right : bar graph depicts the Pearson’s coefficient of colocalization in control and SCD mouse liver. The error bars represent SD. Arrowheads indicate liver sinusoidal endothelial cells. * P < 0.05, ** P < 0.01. Scale bars, 50 µm.

Article Snippet: CD163 activity was blocked with Antigenic Blocking Peptide CD163 N-epitope (FabGennix), which was administered subcutaneously at the dose of 1 mg/kg mouse body weight or as mentioned in the text.

Techniques: Immunofluorescence, Staining, Control

Journal: American Journal of Physiology - Cell Physiology

Article Title: Hemoglobin scavenger receptor CD163 as a potential biomarker of hemolysis-induced hepatobiliary injury in sickle cell disease

doi: 10.1152/ajpcell.00386.2023

Figure Lengend Snippet: CD163 blocker competes with hemoglobin (Hb) for binding. A : schematic showing the experimental plan to analyze the effect of CD163 blocker in sickle cell disease (SCD) mouse liver. B : bar graph depicting the results of heme ELISA assay post administration of CD163 blocker in the plasma of SCD mice. C : schematic showing the experimental plan to examine the mode of action of the CD163 blocker used in blocking CD163-Hb binding. HO-1, heme oxygenase-1. D : bar graphs depicting the results of heme ELISA assay post CD163 blocker administration in the plasma of SCD mice. oxHb, oxyhemoglobin; Veh, vehicle. The error bars represent SD. * P < 0.05. Figure created with BioRender.com.

Article Snippet: CD163 activity was blocked with Antigenic Blocking Peptide CD163 N-epitope (FabGennix), which was administered subcutaneously at the dose of 1 mg/kg mouse body weight or as mentioned in the text.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Blocking Assay

Journal: American Journal of Physiology - Cell Physiology

Article Title: Hemoglobin scavenger receptor CD163 as a potential biomarker of hemolysis-induced hepatobiliary injury in sickle cell disease

doi: 10.1152/ajpcell.00386.2023

Figure Lengend Snippet: Heme oxygenase-1 (HO-1) positively regulates CD163 level in sickle cell disease (SCD) mouse liver. A and B : representative immunohistochemistry image ( A ) and colocalization analysis ( B ) show increased colocalization of CD163 and HO-1 in hepatic Kupffer cells of SCD mouse liver. CON, control. C : qRT-PCR analysis of hepatocytes and nonhepatocytes isolated from control mouse liver exhibits increased expression of both CD163 and HO-1 in the nonhepatocyte cell population. D : the representative images acquired during proximity ligation assay (PLA) show increased interactions between CD163 and HO-1 in SCD mice compared to littermate control mice. Scale bars, 50 µm. E : fluorescence intensity quantification of PLA demonstrating significantly higher intensity in the SCD group compared to control mice. FOV, field of view. F and G : series of Western blot micrographs (for the treatment pattern see ) showing the levels of HO-1 in liver lysates of control mice, untreated SCD mice, and SCD mice after treatment with iron dextran ( F ) and treatments with clodronate and hemoglobin as well as undergoing hypoxic conditions ( G ). H , left : representative Western blot micrographs showing decreased levels of CD163 protein expression after HO-1 knockdown in Hep3B human hepatoma cell line compared with untreated cells. Right : densitometric analysis of micrographs shows significantly reduced levels of CD163 protein expression after HO-1 knockdown and confirms the efficiency of performed knockdown in control and SCD mouse liver tissue. * P < 0.05; # P < 0.06–0.08.

Article Snippet: CD163 activity was blocked with Antigenic Blocking Peptide CD163 N-epitope (FabGennix), which was administered subcutaneously at the dose of 1 mg/kg mouse body weight or as mentioned in the text.

Techniques: Immunohistochemistry, Control, Quantitative RT-PCR, Isolation, Expressing, Proximity Ligation Assay, Fluorescence, Western Blot, Knockdown

Journal: American Journal of Physiology - Cell Physiology

Article Title: Hemoglobin scavenger receptor CD163 as a potential biomarker of hemolysis-induced hepatobiliary injury in sickle cell disease

doi: 10.1152/ajpcell.00386.2023

Figure Lengend Snippet: The CD163 and heme oxygenase-1 (HO-1) signaling pathway in sickle cell disease (SCD)-related liver complications. Schematic diagram depicting the HO-1-CD163 interaction in SCD mouse liver. Cell-free hemoglobin (Hb) released after intravascular hemolysis is scavenged by plasma haptoglobin (HP), which chaperones it to the liver for hemoglobin scavenger receptor CD163-dependent clearance by macrophages. Cell-free Hb on its own can also bind to CD163, albeit less efficiently. In both cases, it leads to endocytosis and degradation of Hb and the release of heme. HO-1 metabolizes heme to produce carbon monoxide, iron, and biliverdin. Along with its role in heme degradation, HO-1 also regulates CD163 expression in the hepatic Kupffer cells. Loss of HO-1 in the liver reduces CD163 expression, which can further impact the Hb-heme metabolism in SCD liver. RBC, red blood cell. Figure created with BioRender.com.

Article Snippet: CD163 activity was blocked with Antigenic Blocking Peptide CD163 N-epitope (FabGennix), which was administered subcutaneously at the dose of 1 mg/kg mouse body weight or as mentioned in the text.

Techniques: Clinical Proteomics, Expressing