Journal: Journal of Clinical Medicine

Article Title: Zoledronate/Anti-VEGF Neutralizing Antibody Combination Administration Increases Osteal Macrophages in a Murine Model of MRONJ Stage 0-like Lesions

doi: 10.3390/jcm12051914

Figure Lengend Snippet: Effects of administered drugs on macrophages in the tooth extraction sockets. ( A ) Representative fluorescent image of F4/80 + macrophages (white arrowheads: F4/80 + macrophages, bar: 100 μm). ( B ) The number of F4/80 + macrophages is significantly decreased in Zol/Vab vs. other comparison groups. ( C ) Representative fluorescent images of CD38 + macrophages (white arrowheads: CD38 + macrophages, bar: 100 μm). ( D ) The number of CD38 + macrophages is significantly increased in Zol/Vab vs. other comparison groups. ( E ) Representative fluorescent images of CD163 + macrophages (white arrowheads: CD163 + macrophages, bar: 100 μm). ( F ) The number of CD163 + macrophages is similar among the comparison groups. ( G ) The M1/M2 ratio is slightly higher in Vab and Zol/Vab vs. VC. ( H ) Representative fluorescent images of CD169 + macrophages (osteomacs) (white arrowheads: CD169 + macrophages, bar: 100 μm). ( I ) The number of CD169 + macrophages is greater in Zol/Vab vs. other comparison groups. Graphs show means ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, †† p < 0.01; n = 5 mice/group.

Article Snippet: M1 macrophages were visualized using the cocktail of pan-macrophage marker rat anti-mouse monoclonal antibody to F4/80 at 1:25 (ab16911, Abcam) and mouse anti-human CD38 monoclonal antibody at 1:50 (sc-374650; Santa Cruz Biotechnology, Dallas, TX, USA), followed by the cocktail of the Alexa FluorTM 546 goat anti-rat IgG (H+L) cross-adsorbed secondary antibody at 1:200 (A-11081, Invitrogen) and goat anti-mouse IgG (H&L) (FITC) (ab6785, Abcam) (both 1:200).

Techniques: Extraction, Comparison

Journal: Orthopaedic surgery

Article Title: CD38 Drives Progress of Osteoarthritis by Affecting Cartilage Homeostasis.

doi: 10.1111/os.13258

Figure Lengend Snippet: Fig. 1 CD38 mRNA (A) and protein expression (B) were detected by RT-PCR and immunofluorescence in control and DMM groups. CD38 expression was upregulated during the development of osteoarthritis. Quantitative analysis of the relative density of CD38 by densitometric analysis. Scale

Article Snippet: See the T erm s and C onditions (https://onlinelibrary.w iley.com /term s-and-conditions) on W iley O nline L ibrary for rules of use; O A articles are governed by the applicable C reative C om m ons L icense OK, USA); FBS (26,140,079, Life Technologies, Frederick, MD, USA); Cas9 enzyme (LentiCRISPR v2; Addgene plasmid repository #52961, Watertown, MA, USA); Alcian blue solution (B8438, Sigma-Aldrich, USA); Safranin O (477– 73-6, Sigma-Aldrich, USA); Fast Green (2353-45-9, SigmaAldrich, St. Louis, MO, USA); Primary antibodies used were as follows: anti-CD38 antibody (sc-37,465, Santa Cruz, USA); anti-β-actin (A1978, Sigma-Aldrich, St. Louis, MO, USA), anti-Col2 (SAB4500366-100UG, Sigma-Aldrich, USA) and anti-aggrecan (AB1031, Sigma-Aldrich, St. Louis, MO, USA).Secondary antibodies used were as follows: Goat antirabbit immunoglobulin-G (ab150077, Abcam, Waltham, PA, USA); Fluorescent secondary antibody (926–32,212, LI-COR, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control

Journal: Orthopaedic surgery

Article Title: CD38 Drives Progress of Osteoarthritis by Affecting Cartilage Homeostasis.

doi: 10.1111/os.13258

Figure Lengend Snippet: Fig. 2 CD38 and chondrogenic marker expression during differentiation. (A) Cartilaginous nodules and extracellular matrix formed after 6 days of

Article Snippet: See the T erm s and C onditions (https://onlinelibrary.w iley.com /term s-and-conditions) on W iley O nline L ibrary for rules of use; O A articles are governed by the applicable C reative C om m ons L icense OK, USA); FBS (26,140,079, Life Technologies, Frederick, MD, USA); Cas9 enzyme (LentiCRISPR v2; Addgene plasmid repository #52961, Watertown, MA, USA); Alcian blue solution (B8438, Sigma-Aldrich, USA); Safranin O (477– 73-6, Sigma-Aldrich, USA); Fast Green (2353-45-9, SigmaAldrich, St. Louis, MO, USA); Primary antibodies used were as follows: anti-CD38 antibody (sc-37,465, Santa Cruz, USA); anti-β-actin (A1978, Sigma-Aldrich, St. Louis, MO, USA), anti-Col2 (SAB4500366-100UG, Sigma-Aldrich, USA) and anti-aggrecan (AB1031, Sigma-Aldrich, St. Louis, MO, USA).Secondary antibodies used were as follows: Goat antirabbit immunoglobulin-G (ab150077, Abcam, Waltham, PA, USA); Fluorescent secondary antibody (926–32,212, LI-COR, USA).

Techniques: Marker, Expressing

Journal: Orthopaedic surgery

Article Title: CD38 Drives Progress of Osteoarthritis by Affecting Cartilage Homeostasis.

doi: 10.1111/os.13258

Figure Lengend Snippet: Fig. 3 Ablation of CD38 promote chondrogenic differentiation. (A) 100 nM and 1 μM 78c treatment increases extracellular matrix formation in micromass cultures (n = 3). Alcian blue staining intensity was measured. Scale bar = 1 mm; (B) Compared with 100 nM 78c treatment, 1 μM

Article Snippet: See the T erm s and C onditions (https://onlinelibrary.w iley.com /term s-and-conditions) on W iley O nline L ibrary for rules of use; O A articles are governed by the applicable C reative C om m ons L icense OK, USA); FBS (26,140,079, Life Technologies, Frederick, MD, USA); Cas9 enzyme (LentiCRISPR v2; Addgene plasmid repository #52961, Watertown, MA, USA); Alcian blue solution (B8438, Sigma-Aldrich, USA); Safranin O (477– 73-6, Sigma-Aldrich, USA); Fast Green (2353-45-9, SigmaAldrich, St. Louis, MO, USA); Primary antibodies used were as follows: anti-CD38 antibody (sc-37,465, Santa Cruz, USA); anti-β-actin (A1978, Sigma-Aldrich, St. Louis, MO, USA), anti-Col2 (SAB4500366-100UG, Sigma-Aldrich, USA) and anti-aggrecan (AB1031, Sigma-Aldrich, St. Louis, MO, USA).Secondary antibodies used were as follows: Goat antirabbit immunoglobulin-G (ab150077, Abcam, Waltham, PA, USA); Fluorescent secondary antibody (926–32,212, LI-COR, USA).

Techniques: Staining

Journal: Orthopaedic surgery

Article Title: CD38 Drives Progress of Osteoarthritis by Affecting Cartilage Homeostasis.

doi: 10.1111/os.13258

Figure Lengend Snippet: Fig. 4 CRISPR/Cas9 Lentivirus knockout of CD38 promote chondrogenic differentiation. A. CRISPR/Cas9 Lentivirus transfection effectively

Article Snippet: See the T erm s and C onditions (https://onlinelibrary.w iley.com /term s-and-conditions) on W iley O nline L ibrary for rules of use; O A articles are governed by the applicable C reative C om m ons L icense OK, USA); FBS (26,140,079, Life Technologies, Frederick, MD, USA); Cas9 enzyme (LentiCRISPR v2; Addgene plasmid repository #52961, Watertown, MA, USA); Alcian blue solution (B8438, Sigma-Aldrich, USA); Safranin O (477– 73-6, Sigma-Aldrich, USA); Fast Green (2353-45-9, SigmaAldrich, St. Louis, MO, USA); Primary antibodies used were as follows: anti-CD38 antibody (sc-37,465, Santa Cruz, USA); anti-β-actin (A1978, Sigma-Aldrich, St. Louis, MO, USA), anti-Col2 (SAB4500366-100UG, Sigma-Aldrich, USA) and anti-aggrecan (AB1031, Sigma-Aldrich, St. Louis, MO, USA).Secondary antibodies used were as follows: Goat antirabbit immunoglobulin-G (ab150077, Abcam, Waltham, PA, USA); Fluorescent secondary antibody (926–32,212, LI-COR, USA).

Techniques: CRISPR, Knock-Out, Transfection

Journal: Biological Research

Article Title: Aβ promotes CD38 expression in senescent microglia in Alzheimer’s disease

doi: 10.1186/s40659-022-00379-1

Figure Lengend Snippet: Aβ1-40 induced energy metabolism disorder and mitochondrial dysfunction in BV2 cells. Scale bar: 50 μm. A Morphological changes of BV2 cells treated with Aβ1-40 (1 μM). B SA-β-Gal positive stained area. C Representative images of BV2 cells immunolabeled for P16 (red) and P21 (red). Nuclear staining (DAPI) is shown in blue. D Cell viability of BV2 cells E ATP level of BV2 cells. F NAD + level of BV2 cells. G NADH level of BV2 cells. H NAD + /NADH ratio of BV2 cells. I ROS level of BV2 cells. J MMP level of BV2 cells. K CD38 enzymatic activity of BV2 cells. L P16, P21, and CD38 protein expression level of BV2 cells. All data are expressed as the mean ± standard error. **P < 0.01 vs. control group

Article Snippet: Mouse microglia BV2 cells were purchased from the Cell Bank of Chinese Academy of Sciences; Dulbecco’s modified Eagle medium (12100046), fetal bovine serum (10099141C), and penicillin/streptomycin (10378016) were purchased from Gibco; Aβ protein fragment 1–40 (A1075) was purchased from Sigma-Aldrich; Lipofectamine 2000 Transfection Reagent (ab51243) was purchased from Abcam; CD38 siRNA (AM160089) was purchased from Invitrogen; CD38 CRISPR activating plasmid (sc-419548), CD38 antibody (SC374650), and M-IgGκ BP-HRP (sc-516102) were purchased from Santa Cruz Biotechnology; anti-rabbit IgG HRP-linked antibody (5490S) and β-actin mouse monoclonal antibody (3700) were purchased from Cell Signaling Technology; P21 rabbit antibody (AF5252), P16 rabbit antibody (AF1069), Iba1 rabbit antibody (AF7143), Cell Proliferation and Cytotoxicity Assay Kit (C0009), Enhanced ATP Assay Kit (S0027), Reactive Oxygen Species Assay Kit (S0033S), Mitochondrial Membrane Potential Assay Kit (C2006), and Mouse amyloid beta peptide 1–42 ELISA Kit (MU30114) were purchased from Bioswamp; Mouse TNF-α ELISA Kit (PT512), Mouse IL-6 ELISA Kit (PT312), Mouse IL-1β ELISA Kit (PT512), Senescence β-Galactosidase Staining Kit (C0602), Alexa Fluor 488 goat anti-rabbit IgG (a0423), DAPI-staining solution (C1006), phosphate buffered saline (C0221A) and bovine serum albumin (ST025) were purchased from Shanghai Beyotime Biotechnology; and NAD/NADH Quantitation Colorimetric Kit (K337) was purchased from Biovision.

Techniques: Staining, Immunolabeling, Activity Assay, Expressing, Control

Journal: Biological Research

Article Title: Aβ promotes CD38 expression in senescent microglia in Alzheimer’s disease

doi: 10.1186/s40659-022-00379-1

Figure Lengend Snippet: CD38 knockout improves energy dysmetabolism and reduces the expression of inflammatory factors in Aβ1-40 injured BV2 cells. Scale bar: 50 μm. A Representative image of BV2 microglial cells exposed to Aβ1-40 (1 μM), and overexpressed or knockdown CD38 immunolabeled for CD38 (green). Nuclear staining (DAPI) is shown in blue. B CD38 protein expression level of BV2 cells. C NAD + level of BV2 cells. D NADH level of BV2 cells. E NAD + /NADH ratio of BV2 cells. F ATP level of BV2 cells. G ROS level of BV2 cells. H MMP level of BV2 cells. I IL-1β level of BV2 cells. J IL-6 level of BV2 cells. K TNF-α level of BV2 cells. All data are expressed as the mean ± standard error. **P < 0.01 vs. control group; ## P < 0.01 vs. Aβ1–40 group; △△ P < 0.01

Article Snippet: Mouse microglia BV2 cells were purchased from the Cell Bank of Chinese Academy of Sciences; Dulbecco’s modified Eagle medium (12100046), fetal bovine serum (10099141C), and penicillin/streptomycin (10378016) were purchased from Gibco; Aβ protein fragment 1–40 (A1075) was purchased from Sigma-Aldrich; Lipofectamine 2000 Transfection Reagent (ab51243) was purchased from Abcam; CD38 siRNA (AM160089) was purchased from Invitrogen; CD38 CRISPR activating plasmid (sc-419548), CD38 antibody (SC374650), and M-IgGκ BP-HRP (sc-516102) were purchased from Santa Cruz Biotechnology; anti-rabbit IgG HRP-linked antibody (5490S) and β-actin mouse monoclonal antibody (3700) were purchased from Cell Signaling Technology; P21 rabbit antibody (AF5252), P16 rabbit antibody (AF1069), Iba1 rabbit antibody (AF7143), Cell Proliferation and Cytotoxicity Assay Kit (C0009), Enhanced ATP Assay Kit (S0027), Reactive Oxygen Species Assay Kit (S0033S), Mitochondrial Membrane Potential Assay Kit (C2006), and Mouse amyloid beta peptide 1–42 ELISA Kit (MU30114) were purchased from Bioswamp; Mouse TNF-α ELISA Kit (PT512), Mouse IL-6 ELISA Kit (PT312), Mouse IL-1β ELISA Kit (PT512), Senescence β-Galactosidase Staining Kit (C0602), Alexa Fluor 488 goat anti-rabbit IgG (a0423), DAPI-staining solution (C1006), phosphate buffered saline (C0221A) and bovine serum albumin (ST025) were purchased from Shanghai Beyotime Biotechnology; and NAD/NADH Quantitation Colorimetric Kit (K337) was purchased from Biovision.

Techniques: Knock-Out, Expressing, Knockdown, Immunolabeling, Staining, Control

Journal: Biological Research

Article Title: Aβ promotes CD38 expression in senescent microglia in Alzheimer’s disease

doi: 10.1186/s40659-022-00379-1

Figure Lengend Snippet: Inhibition of CD38 expression improves energy metabolism disorder in APP/PS1 mice and reduces the neuroinflammatory response. Scale bar: 50 μm. A ATP level of the hippocampus. B ATP level of the cortex. C NAD + level of the hippocampus. D NADH level of the hippocampus. E NAD + /NADH ratio of the hippocampus. F NAD + level of the cortex. G NADH level of the cortex. H NAD + /NADH ratio of the cortex. I MMP of the hippocampus. J MMP of the cortex. K ROS level of the hippocampus. L ROS level of the cortex. M IL-1β level of the serum. N TNF-α level of the serum. O IL-6 level of the serum. P IL-1β level of the hippocampus. Q TNF-α level of the hippocampus. R IL-6 level of the hippocampus. S IL-6 level of the cortex. T TNF-α level of the cortex. U Representative images of Iba1 + and senile plaque immunolabeling in the hippocampus and cortex. *P < 0.05 and **P < 0.01 vs. control group; # P < 0.05 and ## P < 0.01 vs. model group; △ P < 0.05 and △△ P < 0.01

Article Snippet: Mouse microglia BV2 cells were purchased from the Cell Bank of Chinese Academy of Sciences; Dulbecco’s modified Eagle medium (12100046), fetal bovine serum (10099141C), and penicillin/streptomycin (10378016) were purchased from Gibco; Aβ protein fragment 1–40 (A1075) was purchased from Sigma-Aldrich; Lipofectamine 2000 Transfection Reagent (ab51243) was purchased from Abcam; CD38 siRNA (AM160089) was purchased from Invitrogen; CD38 CRISPR activating plasmid (sc-419548), CD38 antibody (SC374650), and M-IgGκ BP-HRP (sc-516102) were purchased from Santa Cruz Biotechnology; anti-rabbit IgG HRP-linked antibody (5490S) and β-actin mouse monoclonal antibody (3700) were purchased from Cell Signaling Technology; P21 rabbit antibody (AF5252), P16 rabbit antibody (AF1069), Iba1 rabbit antibody (AF7143), Cell Proliferation and Cytotoxicity Assay Kit (C0009), Enhanced ATP Assay Kit (S0027), Reactive Oxygen Species Assay Kit (S0033S), Mitochondrial Membrane Potential Assay Kit (C2006), and Mouse amyloid beta peptide 1–42 ELISA Kit (MU30114) were purchased from Bioswamp; Mouse TNF-α ELISA Kit (PT512), Mouse IL-6 ELISA Kit (PT312), Mouse IL-1β ELISA Kit (PT512), Senescence β-Galactosidase Staining Kit (C0602), Alexa Fluor 488 goat anti-rabbit IgG (a0423), DAPI-staining solution (C1006), phosphate buffered saline (C0221A) and bovine serum albumin (ST025) were purchased from Shanghai Beyotime Biotechnology; and NAD/NADH Quantitation Colorimetric Kit (K337) was purchased from Biovision.

Techniques: Inhibition, Expressing, Immunolabeling, Control

Journal: Biological Research

Article Title: Aβ promotes CD38 expression in senescent microglia in Alzheimer’s disease

doi: 10.1186/s40659-022-00379-1

Figure Lengend Snippet: Inhibition of CD38 expression improves the cognitive learning ability of APP/PS1 mice. A Escape latency. B Total time. C Total distance. D Platform crossings. E Swimming trajectory. F New object recognition index. G New object recognition trajectory. H Aβ1-42 concentration of the hippocampus. I Aβ1-42 concentration of the cortex. J Aβ1-42 concentration of the serum. *P < 0.05 and **P < 0.01 vs. control group; # P < 0.05 and ## P < 0.01 vs. model group; △ P < 0.05 and △△ P < 0.01

Article Snippet: Mouse microglia BV2 cells were purchased from the Cell Bank of Chinese Academy of Sciences; Dulbecco’s modified Eagle medium (12100046), fetal bovine serum (10099141C), and penicillin/streptomycin (10378016) were purchased from Gibco; Aβ protein fragment 1–40 (A1075) was purchased from Sigma-Aldrich; Lipofectamine 2000 Transfection Reagent (ab51243) was purchased from Abcam; CD38 siRNA (AM160089) was purchased from Invitrogen; CD38 CRISPR activating plasmid (sc-419548), CD38 antibody (SC374650), and M-IgGκ BP-HRP (sc-516102) were purchased from Santa Cruz Biotechnology; anti-rabbit IgG HRP-linked antibody (5490S) and β-actin mouse monoclonal antibody (3700) were purchased from Cell Signaling Technology; P21 rabbit antibody (AF5252), P16 rabbit antibody (AF1069), Iba1 rabbit antibody (AF7143), Cell Proliferation and Cytotoxicity Assay Kit (C0009), Enhanced ATP Assay Kit (S0027), Reactive Oxygen Species Assay Kit (S0033S), Mitochondrial Membrane Potential Assay Kit (C2006), and Mouse amyloid beta peptide 1–42 ELISA Kit (MU30114) were purchased from Bioswamp; Mouse TNF-α ELISA Kit (PT512), Mouse IL-6 ELISA Kit (PT312), Mouse IL-1β ELISA Kit (PT512), Senescence β-Galactosidase Staining Kit (C0602), Alexa Fluor 488 goat anti-rabbit IgG (a0423), DAPI-staining solution (C1006), phosphate buffered saline (C0221A) and bovine serum albumin (ST025) were purchased from Shanghai Beyotime Biotechnology; and NAD/NADH Quantitation Colorimetric Kit (K337) was purchased from Biovision.

Techniques: Inhibition, Expressing, Concentration Assay, Control