primary antibodies directed against neun  (Millipore)

Journal: Cell Death & Disease

Article Title: PHGDH-mediated serine synthesis in astrocytes supports neuroinflammation by sustaining NADH level to promote histone acetylation

doi: 10.1038/s41419-025-07732-8

Figure Lengend Snippet: A , B The immunofluorescence staining of PHGDH, GFAP, Iba-1, NeuN and DAPI in brain tissue of C57BL/6 mice before and after LPS stimulation and the statistics of colocalization calculated as Pearson’s correlation coefficient, r. Scale, 100 μm, n = 7. C immunofluorescence staining representation of PHGDH, GFAP and Iba-1 in substantia nigra of brain tissue of C57BL/6 mice induced by MPTP. Scale, 100 μm, n = 4. D Statistics of relative PHGDH fluorescence intensity per astrocyte. 54 cells from 6 mice. Statistics of relative PHGDH fluorescence intensity per microglia. 45 cells from 4 mice. The data are means ± SD, for all panels: * P < 0.05, ** P < 0.01, *** P < 0.001, n.s. no significance by two-way ANOVA analysis followed by Bonferroni Test ( B ) and one-way ANOVA analysis followed by Student’s t -test ( D ). All data are representative of or combined from at least three independent experiments.

Article Snippet: Brain tissues were fixed in 4% PFA in phosphate buffer for 12 h at room temperature (RT) and incubated at 4 °C in phosphate buffer containing 30% sucrose for 48 h. The cryosections were permeabilized and blocked in PBS containing 0.3% triton and 10% BSA for 2 h and incubated at 4 °C overnight with the primary antibody against PHGDH (1:1000), GFAP (1:2000, DAKO), Iba1(1:1000, Wako), NeuN (1:1000, Millipore), P-P65(1:1000, CST 3033).

Techniques: Immunofluorescence, Staining, Fluorescence

Journal: Cell Death & Disease

Article Title: PHGDH-mediated serine synthesis in astrocytes supports neuroinflammation by sustaining NADH level to promote histone acetylation

doi: 10.1038/s41419-025-07732-8

Figure Lengend Snippet: A Representative images of PHGDH, GFAP and DAPI immunofluorescence staining in primary astrocytes treated with LPS or PBS. Scale bar, 20 μm, n = 3. B Representative immunoblotting of phosphorylation (p-) or total protein of astrocyte lysates treated with DMSO or NCT-503 in the presence or absence of LPS, n = 3. C Representative images of GFAP, P-P65 and DAPI immunofluorescence staining after astrocytes treated with DMSO or NCT-503 and treated PBS or LPS for 1 h. Scale bar, 10 μm, n = 3. The data are means ± SD, for all panels: n.s. no significance by two-way ANOVA analysis followed by Bonferroni Test ( B ).

Article Snippet: Brain tissues were fixed in 4% PFA in phosphate buffer for 12 h at room temperature (RT) and incubated at 4 °C in phosphate buffer containing 30% sucrose for 48 h. The cryosections were permeabilized and blocked in PBS containing 0.3% triton and 10% BSA for 2 h and incubated at 4 °C overnight with the primary antibody against PHGDH (1:1000), GFAP (1:2000, DAKO), Iba1(1:1000, Wako), NeuN (1:1000, Millipore), P-P65(1:1000, CST 3033).

Techniques: Immunofluorescence, Staining, Western Blot, Phospho-proteomics