Journal: Bioactive Materials

Article Title: Immune regulative GelMA&Zn 2+ /Ce 3+ -whitlockite scaffolds with continuous ions release for bone regeneration

doi: 10.1016/j.bioactmat.2025.11.009

Figure Lengend Snippet: Evaluation of immune inflammation (RAW264.7) in the scaffolds. A, B) Immunofluorescent staining results and quantitative fluorescence intensity of CD206 in RAW264.7 cells co-cultured with different sample extracts, with CD206 (M2 marker) shown in red (scale bar = 50 μm). C, D) Immunofluorescent staining results and quantitative fluorescence intensity of CD86 in RAW264.7 cells co-cultured with different sample extracts, with CD86 (M1 marker) shown in green (scale bar = 50 μm). E, F) Immunofluorescent staining results and quantitative fluorescence intensity of CD163 in RAW264.7 cells co-cultured with different sample extracts, with CD163 (M2 marker) shown in green (scale bar = 50 μm). G) Flow cytometry analysis of CD86 and CD11b positive cells in inflammatory RAW264.7 cells. H) Fluorescence enzyme labeling for the quantitative analysis of ROS content in macrophages. I) Cell viability of RAW264.7 cells co-cultured with different sample extracts for 1, 3, and 5 days. J) NO content in RAW264.7 cells co-cultured with different sample extracts. K) DCFH-DA fluorescent probe detection to evaluate ROS content in macrophages after stimulation with different sample extracts (scale bar = 100 μm). P values were determined using one-way ANOVA (∗∗∗∗P < 0.0001, ∗∗∗P < 0.001, ∗∗P < 0.01, ∗P < 0.05, N.S., not significant).

Article Snippet: Following stimulation, the cells were blocked with 5 % bovine serum albumin (BSA) for 2 h at room temperature, washed three times with PBS, and incubated overnight at 4 °C with a CD86 polyclonal antibody (1:600, 13395-1-AP, Proteintech, USA) and CD163 polyclonal antibody (1:600, 16646-1-AP, Proteintech, USA).

Techniques: Staining, Fluorescence, Cell Culture, Marker, Flow Cytometry, Labeling

Journal: Bioactive Materials

Article Title: Immune regulative GelMA&Zn 2+ /Ce 3+ -whitlockite scaffolds with continuous ions release for bone regeneration

doi: 10.1016/j.bioactmat.2025.11.009

Figure Lengend Snippet: Evaluation of immune inflammation (BMDM) in the scaffolds. A, B) Immunofluorescent staining results and quantitative fluorescence intensity of CD86 in BMDM cells co-cultured with different sample extracts, with CD86 (M1 marker) shown in green (scale bar = 50 μm). C, D) Immunofluorescent staining results and quantitative fluorescence intensity of CD163 in BMDM cells co-cultured with different sample extracts, with CD163 (M2 marker) shown in green (scale bar = 50 μm). P values were determined using one-way ANOVA (∗∗∗∗P < 0.0001, ∗∗∗P < 0.001, ∗∗P < 0.01, ∗P < 0.05, N.S., not significant).

Article Snippet: Following stimulation, the cells were blocked with 5 % bovine serum albumin (BSA) for 2 h at room temperature, washed three times with PBS, and incubated overnight at 4 °C with a CD86 polyclonal antibody (1:600, 13395-1-AP, Proteintech, USA) and CD163 polyclonal antibody (1:600, 16646-1-AP, Proteintech, USA).

Techniques: Staining, Fluorescence, Cell Culture, Marker

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