Journal: The Journal of Biological Chemistry

Article Title: Effect of microglial Pd1 on glial scar formation after spinal cord injury in mice

doi: 10.1016/j.jbc.2025.108489

Figure Lengend Snippet: Cytokine array analysis revealed Pd1-dependent cytokines and chemokines in LPS-treated microglia . A , qPCR showing the decreased Pd1 mRNA in the microglia after Pd1 siRNA (20 nM and 50 nM) treatment for 48 h. All data are presented as mean ± SD. Statistical significance was indicated by ∗∗ p < 0.01, ∗∗∗ p < 0.001, as determined by one-way ANOVA. A : ANOVA, F (2, 12) = 15.32, p = 0.0005; NC siRNA versus Pd1 siRNA 20 nM, p = 0.0022; NC siRNA versus . Pd1 siRNA 50 nM, p = 0.0004; n = 5 wells per group). B , array membranes of protein expression among NC siRNA, NC siRNA + LPS (5 μg/ml), Pd1 siRNA (50 nM), and Pd1 siRNA (50 nM) + LPS (5 μg/ml) groups in culture medium for microglia. C , the expression of one colony-stimulating factor (G-CSF), one intercellular cell adhesion molecule (CD54), two inflammatory factors (IL-6 and TNF-α), and four chemokines (CXCL10, CCL12, CXCL9, and CCL5) significantly increased following LPS treatment, while the expression of one chemokine (CCL4) decreased. A significant increase in the expression of one inflammatory factor (TNF-α) and one chemokine (CXCL9) after treatment with Pd1 siRNA + LPS. All data are presented as mean ± SD. Statistical significance was indicated by ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, as determined by two-way ANOVA. (Figure 6C: two-way ANOVA; G-CSF, NC siRNA versus Pd1 siRNA + LPS, p = 0.0016, Pd1 siRNA versus Pd1 siRNA + LPS, p = 0.0021; CD54, NC siRNA versus Pd1 siRNA, p = 0.0245, NC siRNA versus Pd1 siRNA + LPS, p = 0.0185; IL-6, NC siRNA versus NC siRNA + LPS, p < 0.0001, NC siRNA versus Pd1 siRNA + LPS, p < 0.0001, NC siRNA + LPS versus Pd1 siRNA, p < 0.0001; Pd1 siRNA versus Pd1 siRNA + LPS, p < 0.0001; CXCL10, NC siRNA versus NC siRNA + LPS, p < 0.0001, NC siRNA versus Pd1 siRNA, p < 0.0001, NC siRNA versus Pd1 siRNA + LPS, p < 0.0001, Pd1 siRNA versus Pd1 siRNA + LPS, p = 0.0481; CCL12, NC siRNA versus NC siRNA + LPS, p = 0.0012, NC siRNA versus Pd1 siRNA + LPS, p = 0.0017, NC siRNA + LPS versus Pd1 siRNA, p = 0.0012; Pd1 siRNA versus Pd1 siRNA + LPS, p = 0.0017; CXCL9, NC siRNA versus Pd1 siRNA + LPS, p < 0.0001, NC siRNA + LPS versus Pd1 siRNA + LPS, p = 0.0156, Pd1 siRNA versus Pd1 siRNA + LPS, p = 0.0145; CCL4, NC siRNA versus NC siRNA + LPS, p < 0.0001, NC siRNA versus Pd1 siRNA, p = 0.0062, NC siRNA versus Pd1 siRNA + LPS, p < 0.0001, Pd1 siRNA versus Pd1 siRNA + LPS, p = 0.0143; CXCL2, NC siRNA versus NC siRNA + LPS, p = 0.0121, NC siRNA versus Pd1 siRNA, p < 0.0001, NC siRNA versus Pd1 siRNA + LPS, p = 0.0017; CCL5, NC siRNA versus NC siRNA + LPS, p < 0.0001, NC siRNA versus Pd1 siRNA + LPS, p < 0.0001, NC siRNA + LPS versus Pd1 siRNA, p < 0.0001, Pd1 siRNA versus Pd1 siRNA + LPS, p < 0.0001; TNF-α, NC siRNA versus NC siRNA + LPS, p < 0.0001, NC siRNA versus Pd1 siRNA + LPS, p < 0.0001, NC siRNA + LPS versus Pd1 siRNA, p < 0.0001, NC siRNA + LPS versus Pd1 siRNA + LPS, p = 0.0442, Pd1 siRNA versus Pd1 siRNA + LPS, p < 0.0001; n = 3–4 separate experiments per group). G-CSF, granulocyte colony-stimulating factor; IL-6, interleukin-6; LPS, lipopolysaccharide; NC, negative control; Pd1, programmed cell death protein 1; qPCR, quantitative real-time PCR; siRNA, small interfering RNA; TNF-α, tumor necrosis factor-alpha.

Article Snippet: The levels of specific cytokines secreted by microglia were quantified using a mouse cytokine microarray kit (R&D Systems, ARY006).

Techniques: Expressing, Negative Control, Real-time Polymerase Chain Reaction, Small Interfering RNA

Journal: eLife

Article Title: The role of microglia and their CX3CR1 signaling in adult neurogenesis in the olfactory bulb

doi: 10.7554/elife.30809

Figure Lengend Snippet: Figure 9. Cytokine and Chemokine expression in the OB and serum of WT, Cx3cr1/ and PLX5622-treated mice. (a) Cytokine and Chemokine microarray assay performed on total tissue lysates of OBs from WT, Cx3cr1 /, and PLX5622-terated mice. Expression levels are presented as arbitrary units, measured by densitometry. Using the Bonferroni procedure to correct for multiple comparisons, no significant changes were found between the expressed cytokines. n = 5, pooled in each group. Two-sample t-test. (b) Cytokine and Chemokine microarray assay performed on serum from WT, Cx3cr1/, and PLX5622-terated mice. Cytokine and Chemokine expression levels were measured by densitometry, and are presented as arbitrary units. Using the Bonferroni procedure to correct for multiple comparisons, significant changes were found in the following molecules: (C5a: WT vs PLX, t(2) =12.1, *p=0.0067; CXCL12: WT vs KO, t(2)=11.7, *p=0.007; TIMP1: WT vs PLX, t(2)=8.1, *p=0.014; ICAM1: WT vs KO, t(2)=10.4, *p=0.009). n = 5 pooled in each group. Two-sample t-test. DOI: https://doi.org/10.7554/eLife.30809.021 The following source data is available for figure 9:

Article Snippet: Shaker, Room temp recombinant DNA reagent Details for AAV1 Upenn, PA, U.S.A. Upenn viral core Cat. No. AV-1-PV3365 AAV1 under the CAG promoter, expressing TdTomato commercial assay or kit Cytokine and chemokine microarray assay R and D Systems, MN, U.S.A. Proteome Profiler Mouse Cytokine Array Kit (ARY006) c chemical compound, drug PLX5622 PLEXXIKON Inc., CA, U.S.A. AIN-76A Rodent Diet with PLX5622 1,200 mg PLX5622 (Free Base)/kg chemical compound, drug Control diet PLEXXIKON Inc., CA, U.S.A. AIN-76A Rodent Diet chemical compound, drug BrdU Sigma-Aldrich, MO, U.S.A. Sigma-Aldrich Cat. NO. B5002 10 mg/ml IP injection software, algorithm ImageJ/Figi University of Wisconsin, Madison, WI, U.S.A. (http://rsb.info.nih.gov/ij/), (http://fiji.sc/Fiji) (RRID:SCR_003070) software, algorithm Matlab Mathworks Inc., U.S.A. Matlab, R2014a (RRID:SCR_001622) software, algorithm SPSS I.B.M., U.S.A. SPSS, Version 19 (RRID:SCR_002865)

Techniques: Expressing, Microarray