Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Intracellular Osteopontin of Macrophages Promotes Carotid Plaques Formation by Inducing Foam Cells and Releasing Proinflammatory Cytokines

doi: 10.1161/JAHA.125.041336

Figure Lengend Snippet: A , Western blot analysis on the expression of OPN, CD36, SR‐A, and ABCG1 in macrophages treated with 1% O 2 or 50 μg/mL ox‐LDL for 48 hours. B , Oil Red O staining on foam cells formation at 48 hours after treatments. C , Western blot analysis on the expression of OPN, CD36, SR‐A, and ABCG1 in macrophages with SPP1 knockdown under hypoxia and ox‐LDL stimulations. D , Oil Red O staining on foam cells formation in macrophages with SPP1 knockdown under hypoxia and ox‐LDL stimulations. All data are presented as mean±SD from 4 independent experiments and analyzed with 2‐way ANOVA followed by the Tukey's multiple comparisons test. * P <0.05. ABCG1 indicates ATP‐binding cassette subfamily G member 1; OPN, osteopontin; and ox‐LDL, oxidized low‐density lipoprotein.

Article Snippet: To induce foam cell formation, macrophages were treated with 50 μg/mL ox‐LDL (Athens Research And Technology, GA, USA).

Techniques: Western Blot, Expressing, Staining, Knockdown, Binding Assay

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Intracellular Osteopontin of Macrophages Promotes Carotid Plaques Formation by Inducing Foam Cells and Releasing Proinflammatory Cytokines

doi: 10.1161/JAHA.125.041336

Figure Lengend Snippet: A , Protein expression levels of p‐MLC, MLC, ROCK1, and RhoA in macrophages with SPP1 knockdown under hypoxia and ox‐LDL stimulation. B , Western blot analysis was performed to detect osteopontin in medium, and ponceau S staining was done for total protein normalization. C , The protein expression of OPN, p‐MLC, MLC, ROCK1, and RhoA were examined in macrophages treated with OPN antibody for 48 hours, with SPP1 knockdown under hypoxia and ox‐LDL stimulation. D, E , immunofluorescence staining of CD36, SR‐A, and phalloidin in macrophages with SPP1 knockdown under hypoxia and ox‐LDL stimulation. Scale bars: 20 μm. All data are presented as mean±SD from 4 independent experiments and analyzed with 2‐way ANOVA followed by the Tukey's multiple comparisons test. * P <0.05. CM indicates conditioned medium; MLC, myosin light chain; OPN, osteopontin; ox‐LDL, oxidized low‐density lipoprotein; p‐MLC phosphorylated myosin light chain; RhoA, Ras homolog family member A; and ROCK1, Rho associated coiled‐coil containing protein kinase 1.

Article Snippet: To induce foam cell formation, macrophages were treated with 50 μg/mL ox‐LDL (Athens Research And Technology, GA, USA).

Techniques: Expressing, Knockdown, Western Blot, Staining, Immunofluorescence

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Intracellular Osteopontin of Macrophages Promotes Carotid Plaques Formation by Inducing Foam Cells and Releasing Proinflammatory Cytokines

doi: 10.1161/JAHA.125.041336

Figure Lengend Snippet: A , Analysis of protein expression levels of p‐IKKα, IKKa, p‐IκBα, IκBα, p‐p65, and p65 (as well as concentrations of IL‐6, TNF‐α, and CCL2 in conditioned media ( B )) in macrophages with SPP1 knockdown under hypoxia and ox‐LDL stimulation. C , CM collected from macrophages under different treatment and cocultured with VSMCs for 24 hours. Western blot analysis of MMP9, collagen I, and α‐SMA in VSMCs. D , Immunofluorescence staining for Ki‐67 (green: Ki‐67, blue: DAPI, scale bars: 100μm) and quantification of Ki‐67‐positive cells in VSMCs. All data are presented as mean±SD from 4 independent experiments and analyzed with 2‐way ANOVA followed by Tukey's multiple comparisons test. * P <0.05. α‐SMA indicates alpha smooth muscle actin; CCL2, C‐C motif ligand 2; CM, conditioned media; IL‐6, interleukin‐6; MMP9, matrix metalloproteinase 9; NF‐κB, nuclear factor kappa B; ox‐LDL, oxidized low‐density lipoprotein; p‐IKKα, phosphorylated IκB kinase alpha; p‐IκBα, phosphorylated inhibitor of nuclear factor kappa B alpha; p‐p65, phosphorylated p65; TNF‐α, tumor necrosis factor alpha; and VSMC, vascular smooth muscle cells.

Article Snippet: To induce foam cell formation, macrophages were treated with 50 μg/mL ox‐LDL (Athens Research And Technology, GA, USA).

Techniques: Expressing, Knockdown, Western Blot, Immunofluorescence, Staining

Journal: Experimental and Therapeutic Medicine

Article Title: P2X7R antagonist protects against renal injury in mice with adriamycin nephropathy

doi: 10.3892/etm.2021.11084

Figure Lengend Snippet: Effect of A438079 on the expression of P2X7R, ox-LDL, CXCL16, Bax, caspase-3 and NLRP3 in the glomeruli. Expression of (A and G) P2X7R, (B and H) ox-LDL, (C and I) CXCL16, (D and J) Bax, (E and K) caspase-3 and (F and L) NLRP3 in glomeruli with different groups. ▲ P<0.01 vs. NC group; # P<0.05 and ## P<0.01 vs. ADR group. ADR, adriamycin; A100/200/300, A438079 100/200/300 µmol/kg; CXCL16, C-X-C motif chemokine ligand 16; NC, negative control; ox-LDL, oxidized low density lipoprotein.

Article Snippet: After blocking the sections with 10% goat serum (Beijing Solarbio Science & Technology Co., Ltd.) for 1 h at 37˚C, the sections were incubated with rabbit anti-mouse P2X7R antibody (pAb; 1:300; cat. no. ab109054; Abcam), C-X-C motif chemokine ligand 16 (CXCL16; 1:100; cat. no. bs-1441R; BIOSS), ox-LDL (1:100; cat. no. bsm-1698M; BIOSS), Bax (1:200; cat. no. bsm-52316R; BIOSS), caspase-3 (1:200; cat. no. bsm-33277M; BIOSS) and NLRP3 (1:200; cat. no. ab270449; Abcam) at 4˚C overnight, followed by incubation with HRP-conjugated secondary goat anti-rabbit (1:500; cat. no. ab6721; Abcam)/mouse (1:500; cat. no. ab6789; Abcam) antibody at 37˚C for 30 min.

Techniques: Expressing, Negative Control