beta Tubulin Antibody - #DF7967

Figure 6 IL4I1 is expressed in M2-like macrophages in glioma. (A) Summary of IL4I1 expression in 12 distinct single-cell datasets from glioma patients. (B) Association between IL4I1 and macrophages on XCELL, TIMER, and EPIC algorithms. (C) Association between IL4I1 expression and markers of M1 and M2 macrophages in CGGA and TCGA databases. Color depth and digital scale represent the strength of association. (D) Representative colocalization images from IF staining between IL4I1 and CD206 in clinical glioma specimens. DAPI (blue), IL4I1 (red) and CD206 (green). Scale bar: 20 μm. (E) Measurement of protein and mRNA expression levels for markers (CD11B, CD204, CD86, CD206, and CD163) of THP-1, M0, M1, and M2 macrophages using WB and RT-qPCR. (F) Assessment of IL4I1 expression in various macrophage subtypes (M0, M1, and M2) and glioma cells (U87, LN229, and U251). (G) Representative IF pictures of the difference of IL4I1 among M0, M1, and M2 macrophages, U87, and LN229 cells with DAPI (blue) and IL4I1 (red). *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Figure 5. Ablation of microglia with PLX5622 protects against increased microglial phagocytosis and formation of fear memory. (A) Time course for PLX5622 and AIN-76A administrations and behavior tests. (B) Schematic diagram showing PLX5622 efficacy. (C) Representative confocal images of Iba1 immunostaining (green) of prelimbic microglia of mice with PLX5622 and AIN-76A feeding (scale bars = 100 μm). (D, E) Quantification of Iba1+ cells and Iba1+ area (n = 3 mice, 2 slices per mouse, ***P < 0.001). (F) Percentage of freezing in mice with control or PLX5622 feeding during fear conditioning (n =9 mice, *P < 0.05). (G, H) Percentage of freezing in control or PLX5622 feeding mice during contextual (G) and cued (H) fear memory test (n =9 mice, *P < 0.05, **P < 0.01). (I) Representative images of prelimbic c-Fos (red) and CaMKII (green) in mice with PLX5622 and AIN-76A feeding (scale bars = 100 μm [first row]; 20 μm [the other rows]). (J) Number of prelimbic CaMKII and c-Fos double-labeled neurons (n = 3 mice, 2 slices per mouse, *P < 0.05). (K, L) Protein level of prelimbic SYP and PSD95 detected by WB, and quantitative results for mice treated with or without PLX5622 feeding (n = 6 mice, ***P < 0.001). (M) Representative prelimbic neuron dendritic spine density, and dendritic spine density quantification for mice treated with or without PLX5622 feeding (n = 6 mice, 3 dendritic segments per mouse, **P < 0.01, scale bars = 10 μm).

FIGURE 8 H9c2 cells were treated with 1 μM AngII, (0.6, 1.2 mM) hirudin and 10 μM losartan for 24 h, respectively. (A) The expression of STAT3, MAPK1 and IL-6 were analyzed by qRT-PCR, n = 6. (B) The IL-6 in cell supernatant was assayed by ELISA, n = 5. (C) Western blot for the expression of proteins including STAT3, p-STAT3, MAPK1, p-MAPK1 and IL-6. (D) Quantified by western blot analysis and normalized to control, n = 3. Bars represent the mean ± SD, *p < 0.05, **p < 0.01.