mouse anti aif  (Proteintech)

Journal: Journal of Neuroinflammation

Article Title: iPLA2β loss leads to age-related cognitive decline and neuroinflammation by disrupting neuronal mitophagy

doi: 10.1186/s12974-024-03219-z

Figure Lengend Snippet: Overexpression of iPLA2β in PFC improves cognitive function of old mice. A Scheme of the experimental design. AAV injections were administered 21 days prior to formal behavioral testing. Behavioral test training sessions were conducted two days before the formal testing began. Samples were collected on the seventh day following the initiation of the formal behavioral experiment. B Protein levels of iPLA2β in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups, as assessed by Western blotting and densitometry. C Representative immunofluorescence images of iPLA2β (red) and NeuN (green) in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups. Scale bar: 20 μm. D Representative SA-β-gal staining images in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups. Scale bar: 100 μm. E mRNA expression levels of TGFβ, IL-1β, TNF-α, and CCL2 in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups assessed via qPCR. Normalization was conducted relative to GAPDH expression levels. n = 11. F Representative IHC images of Iba1 in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups. Histogram showing quantification of the Iba1(+) area. Scale bar: 50 μm. G Travel time of the mice to reach the platform during the spatial test. n = 20 per group. H Duration spent by mice in the hidden platform quadrants during the probe trial. n = 20. I Frequency of platform crossings by mice during the probe trial. n = 20 per group. J Recognition index during the Novel object recognition test. Recognition index = time spent exploring a novel object/time spent exploring both objects. n = 20. K Discrimination index during the Novel object recognition test. The discrimination index = (time spent on the novel object − time spent on the familiar object)/ time spent on both objects. n = 20 Data are presented as the mean ± SEM; p values were obtained using the Mann-Whitney U test (B, C, D, and F) and the Kruskal-Wallis test followed by Dunn’s multiple comparisons test (E, G, H, I, J, and K). * p < 0.05. **, p < 0.01; ***, p < 0.001

Article Snippet: Then, brain sections were incubated overnight at 4 °C with the following antibodies: mouse polyclonal iPLA2β (sc-376563, 1:100, Santa Cruz), mouse monoclonal Iba1 (#17198, 1:400, Cell Signaling), or rabbit polyclonal GFAP (ab7260, 1:500, Abcam).

Techniques: Over Expression, Control, Western Blot, Immunofluorescence, Staining, Expressing, MANN-WHITNEY

Journal: eLife

Article Title: Fetal liver macrophages contribute to the hematopoietic stem cell niche by controlling granulopoiesis

doi: 10.7554/elife.86493

Figure Lengend Snippet: Figure 3. Macrophage interaction with CD150+ hematopoietic stem cells (HSCs). (A) Immunostaining of E14.5 fetal liver sections with antibodies against F4/80, Iba1, CD150, and Ter119. Scale bar represents 50 µm. (B) Immunostaining of E14.5 fetal liver whole-mounts with antibodies against F4/80, c-Kit, Tim4, Ter119, Iba1, and CD150. The first row visualizes the interaction between EI macrophages surrounded by Ter119+ erythroblasts. The second and last rows highlight the interaction between macrophages and progenitor cells, including CD150+ long-term HSCs (LT-HSCs). The black arrows on the last row indicate the yz and xz dimensions which are shown on the left and bottom sides. Scale bar represents 5 µm. (C) 3D reconstruction of a E14.5 fetal liver whole mount stained with Iba1, CD150, and DAPI (shown as gray dots indicating CD150- cells). Scale bar represents 20 µm (left) or 10 µm (right). Measured distance of CD150+ and CD150− cells to the next Iba1+ macrophage (D) and average distance of cells to Iba1+ macrophages (n = 42 for CD150+ and n = 524 for CD150−) (E). Unpaired Student’s t-test.

Article Snippet: Samples were incubated overnight in a permeabilization solution (PBS, 0.1% Triton- X, 0.1 M glycine, 5% bovine serum albumin (BSA), 5% goat serum) at room temperature and then stained overnight in staining buffer (PBS, 0.1% Triton- X, 1% BSA) at room temperature as follows: DAPI (1:10,000 dilution; Thermo Fisher), Alexa Fluor 555 rabbit anti- mouse Iba1 (1:200 dilution; Cell Signaling, #36618), CD150 unconjugated (1:100 dilution; Invitrogen, clone 9D1).

Techniques: Immunostaining, Staining

Journal: eLife

Article Title: Fetal liver macrophages contribute to the hematopoietic stem cell niche by controlling granulopoiesis

doi: 10.7554/elife.86493

Figure Lengend Snippet: Figure 5. The effect of macrophage depletion on erythropoiesis. (A) Quantification of F4/80+ macrophage cells in the E14.5 fetal liver of Tnfrsf11a+/+; Spi1f/f (WT) and Tnfrsf11aCre/+; Spi1f/f (KO) embryos using flow cytometry. n = 16 for WT, n = 10 for KO. (B) Immunofluorescent staining of macrophages using Iba1 antibody in E14.5 livers. Representative for n = 3. Scale bar represents 30 µm. (C) Quantification of single live and CD45+ cells in the E14.5 fetal liver of WT and KO knockout embryos using flow cytometry. n = 11 for WT, n = 5 for KO. (D) Hematoxylin and eosin stain (HE) of WT and KO fetal

Article Snippet: Samples were incubated overnight in a permeabilization solution (PBS, 0.1% Triton- X, 0.1 M glycine, 5% bovine serum albumin (BSA), 5% goat serum) at room temperature and then stained overnight in staining buffer (PBS, 0.1% Triton- X, 1% BSA) at room temperature as follows: DAPI (1:10,000 dilution; Thermo Fisher), Alexa Fluor 555 rabbit anti- mouse Iba1 (1:200 dilution; Cell Signaling, #36618), CD150 unconjugated (1:100 dilution; Invitrogen, clone 9D1).

Techniques: Flow Cytometry, Staining, Knock-Out, H&E Stain