eye lysates  (Bertin Technologies)

Journal: Cell reports

Article Title: APOE4-promoted gliosis and degeneration in tauopathy are ameliorated by pharmacological inhibition of HMGB1 release.

doi: 10.1016/j.celrep.2023.113252

Figure Lengend Snippet: Figure 1. APOE4 promotes the nucleocytoplasmic translocation of HMGB1 in hippocampal neurons in tauopathy mice (A) Representative images of immunostaining with anti-HMGB1, anti-NeuN, and DAPI in the dentate gyrus of the hippocampus (scale bar, 40 mm). (B) Representative high-magnification images of immunostaining with anti-HMGB1 and DAPI in the dentate gyrus of the hippocampus (scale bar, 10 mm).

Article Snippet: Measurement of HMGB1 levels with sandwich ELISA Biochemically extracted mouse hippocampal tissue lysates or collected hippocampal interstitial fluid (ISF) were run according to the provided manufacturer protocols (mouse HMGB1; (Novus Biologicals).

Techniques: Translocation Assay, Immunostaining

Journal: Cell reports

Article Title: APOE4-promoted gliosis and degeneration in tauopathy are ameliorated by pharmacological inhibition of HMGB1 release.

doi: 10.1016/j.celrep.2023.113252

Figure Lengend Snippet: Figure 3. APOE4 promotes acetylation of HMGB1 and decreases levels of SIRT1 deacetylase, and removal of neuronal APOE4 reduces HMGB1 acetylation and increases SIRT1 levels (A) Representative images of anti-acetyl-HMGB1 (green) and anti-GAPDH (red) western blots in hippocampal tissue lysates. (B) Quantification of acetyl-HMGB1 levels relative to GAPDH. (C and D) Quantification of acetyl-HMGB1 levels relative to GAPDH in the nuclear fraction (C) and cytoplasmic fraction (D).

Article Snippet: Measurement of HMGB1 levels with sandwich ELISA Biochemically extracted mouse hippocampal tissue lysates or collected hippocampal interstitial fluid (ISF) were run according to the provided manufacturer protocols (mouse HMGB1; (Novus Biologicals).

Techniques: Histone Deacetylase Assay, Western Blot

Journal: Cell reports

Article Title: APOE4-promoted gliosis and degeneration in tauopathy are ameliorated by pharmacological inhibition of HMGB1 release.

doi: 10.1016/j.celrep.2023.113252

Figure Lengend Snippet: Figure 4. Short-term treatment of PS19-E4 mice with HMGB1 inhibitors blocks the nucleocytoplasmic translocation of HMGB1 in hippo- campal neurons and its release into the ISF (A) Experimental design of the HMGB1 inhibitor study, with EP + GA treatment at three doses per week. (B) Representative high-magnification images of immunostaining with anti-HMGB1 and DAPI in hippocampal neurons (scale bar, 10 mm). (C and D) Quantification of the nuclear integrated density (C) and extranuclear integrated density (D) of HMGB1 immunostaining in hippocampal neurons. (E) HMGB1 protein levels measured by ELISA in the hippocampal interstitial fluid (ISF). (F) HMGB1 protein levels measured by ELISA in each collected ISF fraction. Fractions 1 and 2 were excluded from analyses in (E) and (F). (G) Representative images of microglia immunostaining with anti-Iba1 in the hippocampus (scale bar, 500 mm). (H) Quantification of the percentage of Iba1 coverage area in the hippocampus. (I) Representative images of astrocyte immunostaining with anti-GFAP in the hippocampus (scale bar, 500 mm). (J) Quantification of the percentage of GFAP coverage area in the hippocampus. (K) Representative images of activated microglia immunostaining with anti-CD68 in the hippocampus (scale bar, 500 mm). (L) Quantification of the percentage of CD68 coverage area in the hippocampus. (M) Representative images of activated astrocyte immunostaining with anti-S100b in the hippocampus (scale bar, 500 mm). (N) Quantification of the percentage of S100b coverage area in the hippocampus. (legend continued on next page)

Article Snippet: Measurement of HMGB1 levels with sandwich ELISA Biochemically extracted mouse hippocampal tissue lysates or collected hippocampal interstitial fluid (ISF) were run according to the provided manufacturer protocols (mouse HMGB1; (Novus Biologicals).

Techniques: Translocation Assay, Immunostaining, Enzyme-linked Immunosorbent Assay

Journal: Cell reports

Article Title: APOE4-promoted gliosis and degeneration in tauopathy are ameliorated by pharmacological inhibition of HMGB1 release.

doi: 10.1016/j.celrep.2023.113252

Figure Lengend Snippet: Figure 6. Long-term treatment of PS19-E4 mice with HMGB1 inhibitors drastically reduces Tau pathology, myelin deficits, and hippocampal degeneration (A) Representative images of p-Tau immunostaining with AT8 monoclonal antibody in the hippocampus (scale bar, 500 mm). (B) Quantification of the percentage of p-Tau (AT8+) coverage area in the hippocampus. (C) Representative images of myelin sheath staining with anti-MBP and DAPI in the stratum radiatum of CA1 in the hippocampus (scale bar, 100 mm). (D) Quantification of the percentage of MBP coverage area in the stratum radiatum of CA1. (E) Representative images of the ventral hippocampus after staining with Sudan black (scale bar, 1 mm). (F and G) Quantification of hippocampal volume (F) and posterior lateral ventricle volume (G). For all representative images and quantified data, mice were 9.5 months of age and belonged to either the PS19-E4 or the PS19-E3 group treated with saline or HMGB1 inhibitors. Quantified data in (B), (D), (F), and (G) (PS19-E4 saline, n = 16; PS19-E4 HMGB1-In, n = 18; PS19-E3 saline, n = 16, PS19-E3 HMGB1-In, n = 14) are represented as the mean ± SEM, one-way ANOVA with Tukey’s post hoc multiple comparisons test. HMGB1-In, HMGB1 inhibitors.

Article Snippet: Measurement of HMGB1 levels with sandwich ELISA Biochemically extracted mouse hippocampal tissue lysates or collected hippocampal interstitial fluid (ISF) were run according to the provided manufacturer protocols (mouse HMGB1; (Novus Biologicals).

Techniques: Immunostaining, Staining, Saline