Journal: Journal of neuroinflammation

Article Title: Brain endothelial CXCL12 attracts protective natural killer cells during ischemic stroke.

doi: 10.1186/s12974-023-02689-x

Figure Lengend Snippet: Fig. 5 NK cells positively regulate behavioral deficits after PT stroke induction. A Coordinates of the lesion on the brain of photothromobotic (PT) mouse model used for beam-walk sensorimotor test analysis. The center of the lesion is 0.3 mm in front of bregma and 2.0 mm lateral to midline, with the 2 mm of diameter. B Nissl staining on the vibratome section of stroke brain at day 2 after PT. Data represent n = 7 mice. C The beam-walk sensorimotor test analysis was performed by calculating percentage of contralateral hindlimb faults. Anti-NK1.1 or IgG2a was injected at day 9 (P-9) and day 2 (P-2) before PT induction and the efficiency of depletion in blood was tested at P-7, P-1, P6, P12 and P18. The videos were recorded just before PT induction (P0) and every second day afterwards. (n = 3 mice for IgG2a, n = 4 mice for anti-NK1.1). D Lesion sizes of brains from the isotype control (IgG2a) (blue) vs. anti-NK1.1 (red) injected mice at P2 and P18 (n = 3 mice for IgG2a, n = 4 mice for anti-NK1.1). E Control (n = 4) and KO (n = 4) mice after PT induction by calculating the percentage of contralateral hindlimb faults.. P2 **p = 0.0058, P4 **P = 0.0053, P6 **P = 0.0096

Article Snippet: For in vivo depletion of NK cells, mice were injected 100 μg of InVivoPlus anti-mouse Nk1.1 (Clone PK136, Bio X Cell) or InVivoPlus mouse IgG2a (Clone C1.18.4, Bio X Cell) resolved in PBS at P-9 and P-2, respectively.

Techniques: Staining, Injection, Control

Journal: Diabetologia

Article Title: Phenotypically distinct anti-insulin B cells repopulate pancreatic islets after anti-CD20 treatment in NOD mice

doi: 10.1007/s00125-019-04974-y

Figure Lengend Snippet: Anti-insulin B cells repopulate pancreatic islets more rapidly than insulin-negative B cells after anti-CD20 treatment. Groups of VH125.hCD20/NOD mice, aged 6–8 weeks old, were injected with 2H7 anti-CD20 or IgG isotype control. Spleen, PLNs and pancreatic islets were analysed for insulin-positive and insulin-negative B cells at 8 weeks and 12 weeks post depletion by flow cytometry. ( a – f ) No. of cells from IgG control-treated (black circles) and 2H7-treated (grey squares) mice for insulin-negative B cells ( a – c ) and insulin-positive B cells ( d – f ) from spleen ( a , d ) PLNs ( b , e ) and islets ( c , f ). ( g – i ) Percentage of B cells repopulated at 8 and 12 weeks after treatment from spleen ( g ), PLNs ( h ) and islets ( i ) of mice shown in ( a – f ). Percentages were calculated as individual numbers from each 2H7-treated mouse / mean number from all IgG control antibody-treated mice. Horizontal lines represent the median value. Data represent three independent experiments. At 8 weeks, n = 7 (spleen), n = 7 (PLNs) and n = 9 (islets) for control IgG-treated mice and n = 10 (spleen), n = 9 (PLNs) and n = 12 (islets) for 2H7-treated mice. At 12 weeks, n = 8 (spleen), n = 6 (PLNs) and n = 7 (islets) for control IgG and n = 11 (spleen), n = 7 (PLNs) and n = 11 (islets) for 2H7-treated mice. * p < 0.05 (one-way ANOVA)

Article Snippet: Female VH125.hCD20/NOD mice, 6–8 weeks of age were chosen at random to receive either anti-hCD20 antibody (clone 2H7; Bio-XCell, West Lebanon, NH, USA) or control IgG2b antibody (clone MPC-11; Bio-XCell [ , , ]), as described previously [ ].

Techniques: Injection, Control, Flow Cytometry

Journal: Diabetologia

Article Title: Phenotypically distinct anti-insulin B cells repopulate pancreatic islets after anti-CD20 treatment in NOD mice

doi: 10.1007/s00125-019-04974-y

Figure Lengend Snippet: CD138 int anti-insulin B cells are enriched in pancreatic islets after anti-CD20 treatment. Groups of 6- to 8-week-old VH125.hCD20/NOD mice were injected with 2H7 anti-CD20 or IgG isotype control. Groups of mice ( n = 2 or 3 per group) were pooled and insulin + B cells from pancreatic islets were analysed for four different populations based on CD138 expression: CD138 − (blue); CD138 int IgM + (orange); CD138 int IgM lo (grey) and CD138 hi IgM lo (red). ( a , b ) Representative flow plots showing gating on live CD3 − CD11b − CD11c − ( a ) and graph showing the overall percentages of the four different populations ( b ). ( c , d ) Representative flow plots showing insulin − CD19 + , insulin + CD19 + and insulin + CD19 − cells ( c ) and graph showing the overall percentages of these cells ( d ); 2H7 (black circles), IgG (grey circles). ( e ) Representative flow plots showing CD138 and IgM expression in insulin + CD19 + and insulin + CD19 − cells. ( f , g ) Graphs showing CD138 and IgM populations on insulin + CD19 + ( f ) and insulin + CD19 − cells ( g ) ( n = 5 groups for control IgG treatment; n = 4 groups for 2H7 treatment). Horizontal lines represent the median values. Data represent two independent experiments. * p < 0.05 (one-way ANOVA)

Article Snippet: Female VH125.hCD20/NOD mice, 6–8 weeks of age were chosen at random to receive either anti-hCD20 antibody (clone 2H7; Bio-XCell, West Lebanon, NH, USA) or control IgG2b antibody (clone MPC-11; Bio-XCell [ , , ]), as described previously [ ].

Techniques: Injection, Control, Expressing

Journal: Matrix biology : journal of the International Society for Matrix Biology

Article Title: Elevated TGFβ signaling contributes to ocular anterior segment dysgenesis in Col4a1 mutant mice

doi: 10.1016/j.matbio.2022.05.001

Figure Lengend Snippet: (A) Schematic illustration of the 1D11 administration paradigm for histological analysis. (B) Representative images of H&E stained sections (left) showing whole eyes (top panels) and corneas (lower panels) from E18.5 mice and quantification graph (right) showing increased corneal stromal thickness in Col4a1+/G1344D mice treated with 1D11 compared to those that received the control IgG1 antibody. Scale bars = 200 μm (top) and 50 μm (bottom). n = 20 and 24 corneas from IgG1- and 1D11-treated Col4a1+/+ mice, and 24 and 30 corneas from IgG1- and 1D11-treated Col4a1+/G1344D mice, respectively. (C) Schematic illustration of the 1D11 administration paradigm for qPCR analyses. (D) qPCR analyses revealed that 1D11 treatment partially prevented the increased expression of TGFβ target genes in anterior segments from P0 Col4a1+/G1344D mice compared to their IgG1-treated counterparts. n = 5–6 samples per genotype. Data are presented as mean ± SD. *p < 0.05; **p < 0.01; ***p < 0.001; and ****p < 0.0001, two-way ANOVA and Tukey’s multiple comparison test.

Article Snippet: Timed-pregnant B6 females crossed with Col4a1 +/ G1344D males were injected intraperitoneally with the 1D11 pan-TGFβ neutralizing antibody (clone 1D11.16.8, BioXCell, West Lebanon, NH) or IgG1 isotype control antibody (clone MOPC-21, BioXCell) diluted in inVivoPure Dilution Buffer (pH 7.0, BioXCell) (20 mg/kg) every other day from E8.5 to E16.5 and animals were harvested at E18.5 or P0 for histological and molecular analyses, respectively.

Techniques: Staining, Control, Expressing, Comparison

Journal: Aging Cell

Article Title: The aged hematopoietic system promotes hippocampal‐dependent cognitive decline

doi: 10.1111/acel.13192

Figure Lengend Snippet: Cyclophilin A regulates cellular and cognitive hallmarks of hippocampal aging. (a) Schematic showing hydrodynamic tail vein injection (HDTVI)‐mediated CyPA overexpression (OE) paradigm in young mice. (b–d) Young CyPA‐OE or GFP control mice were tested in novel object recognition (b; NOR; n = 8–12/group), contextual fear conditioning (c; FC; n = 15–16), and cued FC (d; n = 15–16). (e, f) Representative fields (e; scale bar = 100 μm) and quantification (f) of DCX+ cells ( n = 11/group), in DG of young CyPA‐OE and GFP control mice. (g, h) Representative Western blot (g) and quantification (h) of hippocampal lysates from young CyPA‐OE and GFP control mice, probed with anti‐NR2B, synapsin‐1 (Syn‐1), synaptophysin (Syp), and GAPDH ( n = 5/group). (i) Schematic showing CyPA inhibition paradigm in aged mice. (j–l) Aged mice treated with an anti‐CyPA antibody or IgG isotype control (ctrl) were tested in NOR (j; n = 10–14/group), contextual FC (k; n = 10–14/group), and cued FC (l; n = 10–14/group). (m, n) Representative fields (m; scale bar = 100 μm) and quantification (n) of DCX+ ( n = 8–10/group), in anti‐CyPA or IgG ctrl‐treated old mice. (o, p) Representative Western blot (o) and quantification (p) of hippocampal lysates from anti‐CyPA or IgG ctrl‐treated old mice, probed with anti NR2A, Syn‐1, Syp, and GAPDH ( n = 5/group). All data shown as the mean + SEM . *p < 0.05. t test (c, d, f, h, k, l, n, p). One‐sample t test vs. hypothetical mean of 50 (b, j).

Article Snippet: The anti‐CyPA antibody, 8H7 (Von Ungern‐Sternberg et al., ), or a monoclonal IgG2A isotype control (R&D Systems) was intraperitoneally administered to aged (19 months) mice (20 μg/kg).

Techniques: Injection, Over Expression, Control, Western Blot, Inhibition