Journal: Microcirculation (New York, N.Y. : 1994)

Article Title: Arteriogenesis in murine adipose tissue is contingent on CD68 + /CD206 + macrophages

doi: 10.1111/micc.12341

Figure Lengend Snippet: Fluorescent immunohistochemical labeling reveals an increase in CD68+ and CD206+ cells in inguinal fat pads injected with DiI labeled, M2-polarized macrophages. A.) No fluorescent signal was detectable in inguinal fat pads that were injected with unlabeled cells (scale bar = 50 µm), whereas (B.) DiI labeled cells were visible in inguinal fat pads that were injected with DiI labeled, M2-polarized macrophages (scale bar = 50 µm). C.,D.) Fluorescent immunohistochemical labeling of inguinal fat pad revealed increase in CD68+ cells in (C) DiI+ cell injected tissue when compared to (D) uninjected tissue (scale bar = 20 µm). E.,F.) Fluorescent immunohistochemical labeling of inguinal fat pad revealed increase of CD206+ cells in (E) DiI+ cell injected tissue when compared to (F) uninjected tissue (scale bar = 20 µm) G.) Number of CD68+ cells in injected inguinal fat pads was increased compared to uninjected inguinal fat pads. n = 6 mice. Student’s t-test, * = p-value < 0.05. H.) Number of CD206+ cells was increased in injected inguinal fat pads when compared to uninjected inguinal fat pads. n = 5 mice. Student’s t-test, * = p-value < 0.05. Data are mean ± standard deviation.

Article Snippet: Sample 2 was submerged in 100 microliters of Alexa Fluor 488 conjugated isolectin (1:300, GS-IB 4 , ThermoFisher Scientific, Waltham, MA) and Alexa Fluor 647 anti-mouse CD68 (1:200, Clone FA-11, AbD Serotec, Raleigh, NC) diluted in 5% mouse serum in 0.3% Triton X-100/PBS.

Techniques: Immunohistochemical staining, Labeling, Injection, Standard Deviation

Journal: Microcirculation (New York, N.Y. : 1994)

Article Title: Arteriogenesis in murine adipose tissue is contingent on CD68 + /CD206 + macrophages

doi: 10.1111/micc.12341

Figure Lengend Snippet: Direct injection of clodronate liposomes into the inguinal fat pad significantly depleted local CD68+ macrophage populations after 24 hours. A.,B.) Representative image of control liposome-injected inguinal fat pad (A) and clodronate liposome-injected inguinal fat pad (B) at 6 hours. C.,D.) Representative image of control liposome-injected inguinal fat pad (C) and clodronate liposome-injected inguinal fat pad (D) at 12 hours. E.,F.) Representative image of control liposome-injected inguinal fat pad (E) and clodronate liposome-injected inguinal fat pad (F) at 6 hours. G.) Quantification of CD68+ cells at 6 hours, 12 hours, and 24 hours following either control liposome or clodronate liposome injection. A significant decrease in the number of CD68+ cells was observed 24 hours post-injection in clodronate liposome-injected inguinal fat pads (Student’s t-test, * = p-value < 0.05) relative to control liposome-injected fat pads. Scale bar = 100 µm, n = 3 mice for each time point, 3 unique FOVs for CD68+ cell quantification.

Article Snippet: Sample 2 was submerged in 100 microliters of Alexa Fluor 488 conjugated isolectin (1:300, GS-IB 4 , ThermoFisher Scientific, Waltham, MA) and Alexa Fluor 647 anti-mouse CD68 (1:200, Clone FA-11, AbD Serotec, Raleigh, NC) diluted in 5% mouse serum in 0.3% Triton X-100/PBS.

Techniques: Injection, Liposomes, Control

Journal: Microcirculation (New York, N.Y. : 1994)

Article Title: Arteriogenesis in murine adipose tissue is contingent on CD68 + /CD206 + macrophages

doi: 10.1111/micc.12341

Figure Lengend Snippet: Clodronate liposomes depleted the local macrophage population after ligation surgery and affected the arteriogenic response of collateral vessels. A.) Quantification of CD68+ cells reveals a significant decrease in the number of CD68+ cells within clodronate liposome-treated tissue compared to control liposome-treated tissue. Student’s t-test, ** = p-value < 0.01 B.) Collateral vessel diameters in the clodronate liposome-injected tissues were paired with the diameter measurements from the corresponding collateral vessel branches in the control liposome-injected tissues in the same mouse. Collateral vessels in control liposome-injected fat pads had larger diameters compared to corresponding collateral vessels in clodronate liposome-injected fat pads. Sham and ligated data from a previously published study43 is presented for comparison purposes. Paired t-test, ** = p-value < 0.01. n = 3 mice for all experiments, 4 unique FOVs for CD68+ cell quantification.

Article Snippet: Sample 2 was submerged in 100 microliters of Alexa Fluor 488 conjugated isolectin (1:300, GS-IB 4 , ThermoFisher Scientific, Waltham, MA) and Alexa Fluor 647 anti-mouse CD68 (1:200, Clone FA-11, AbD Serotec, Raleigh, NC) diluted in 5% mouse serum in 0.3% Triton X-100/PBS.

Techniques: Liposomes, Ligation, Control, Injection, Comparison

Journal: Neurobiology of disease

Article Title: Genetic ablation of Gpnmb does not alter synuclein-related pathology.

doi: 10.1016/j.nbd.2021.105494

Figure Lengend Snippet: Fig. 3. Gpnmb knockout does not alter recovery from lysolecithin-induced demyelination. (A) Schematic of lysolecithin demyelination/remyelination model. dpi: days post injection (B) Gpnmb IHC of corpus callosum in wildtype and knockout animals. (C–F) IHC for activated microglia (Cd68), astrocytes (Gfap), degraded myelin (dMBP) and apoptotic cells (cleaved caspase 3). Each point represents the average of up to 3 sections per animal. Integrated intensity averaged over (corpus callosum) area quantified for C-E, number of positive cells per area for F. No significant differences were observed for any of these signals when comparing between knockout and wildtype animals at any of the timepoints (t-test P > 0.05). F′, F′′, zoom in on very rare cleaved caspase 3+ signal, which was not significantly enriched in knockout relative to wildtype and in all cases far below the roughly 100 cells/mm2 previously reported using this assay in cuprizone-treated MerTK knockout mice (Shen et al., 2021). Scale bar in panel C applies as well to D and E. Scale bar in F′ applies as well to F′′.

Article Snippet: Target Figure Vendor Catalog number Link Human GPNMB 2G LS Bio LS-C340379 https://www.lsbio.com/antibodies/gpnmb-antibody-osteoactivin-antibody-clone-oti8a10-ihc-wb-western-ls-c3 40379/350990 Mouse GPNMB 2C-D, 3B, 4B R&D Systems AF2330 https://www.rndsystems.com/products/mouse-osteoactivin-gpnmb-antibody_af2330 Iba1 2G Abcam ab178846 https://www.abcam.com/iba1-antibody-epr16588-ab178846.html Mouse Cd68 2D, 3D, 4B, 4C BioRad MCA1957 https://www.bio-rad-antibodies.com/monoclonal/mouse-cd68-antibody-fa-11-mca1957.html?f=purified Mouse GFAP 2D, 3C, 4B, 4C DAKO Z0334 https://www.agilent.com/en/product/immunohistoch emistry/antibodies-controls/primary-antibodies/glial-fibrillary-acidic-protein-(concentrate)-76683 Mouse dMBP 3E Millipore AB5864 https://www.emdmillipore.com/US/en/product/Anti-Myelin-Basic-Protein-Antibody,MM_NF-AB5864 mouse cleaved caspase 3 3F Cell Signaling 9661S https://www.cellsignal.com/products/primary-antibodies/cleaved-caspase-3-asp175-antibody/9661 p129-αSyn 4B, 4C Wako 010–26,481 https://labchem-wako.fujifilm.com/us/product/detail/W01W0101-2648.html NeuN 4C Millipore MAB377B https://www.emdmillipore.com/US/en/product/Anti-NeuN-Antibody-clone-A60-biotin-conjugated,MM_N F-MAB377B Mouse Th 5C Pel Freez p40101 https://www.pelfreez-bio.com/wp-content/uploads/2014/07/74075-PDS-P40101-Tyrosine-Hydroxylase-Antibod y-Rabbit-Rev-02.pdf Human GPNMB S2A R&D Systems AF2550 https://www.rndsystems.com/products/human-osteoactivin-gpnmb-antibody_af2550 Human TH S2B Millipore AB152 https://www.emdmillipore.com/US/en/product/Anti-Tyrosine-Hydroxylase-Antibody,MM_NF-AB152 Human GPNMB RNA S2B Advanced Cell Diagnostics 413,528 https://acdbio.com/search/site/%252A413528%252A/cms/probes GPNMB knockout Gpnmbtm1.1(KOMP)Vlcg Human SNCA transgenic A53T α-synuclein transgenic line M83 Control Parkinson’s Total number of subjects 10 9 Mean Age at Death (Standard Deviation) 80.1 (12.1) 73 (6.8) Sex 4 female, 5 male, 1 unknown 7 male, 2 unknown R. Brendza et al. Neurobiology of Disease 159 (2021) 105494

Techniques: Knock-Out, Injection