Journal: Journal of Neuroinflammation

Article Title: The protective PLCγ2-P522R variant mitigates Alzheimer’s disease-associated pathologies by enhancing beneficial microglial functions

doi: 10.1186/s12974-025-03387-6

Figure Lengend Snippet: PLCγ2-P522R variant does not increase the plaque-associated APOE or DAM signature in female APP/PS1 mice. A Analysis of total APOE and APOE areas (µm 2 ) within and surrounding (within 0–10- and 10–20-µm distances from the amyloid plaque outline) β-amyloid plaques revealed no differences between the APP/PS1xPLCγ2-P522R (A+/P ki/ki ) and APP/PS1 (A+/P wt/wt ) mice. A+/P wt/wt n = 5, A+/P ki/ki n = 6. Colored disks on the x-axis indicate the analyzed area overlapping (middle circle) and surrounding β-amyloid plaques. The scale bar in the representative immunofluorescence images is 50 μm. Unpaired samples t test. All the data are presented as the means ± SEMs. Each data point represents an individual mouse. B Heatmap of z scored vst-normalized expression of homeostatic as well as stage 1 and stage 2 disease-associated microglia (DAM) signature genes in the bulk temporo-occipital cortex and C ) CD11b+ microglia isolated from the whole brain tissue of 13-month-old wild-type (A-/P wt/wt ), A+/P wt/wt and A+/P ki/ki female mice. Cortex A-/P wt/wt n = 1, A+/P wt/wt n = 4, and A+/P ki/ki n = 4; CD11b+ A-/P wt/wt n = 7, A+/P wt/wt n = 4, and A+/P. ki/ki n = 4

Article Snippet: Soluble and insoluble APOE were measured in lysates using an Apolipoprotein E (APOE) Mouse ELISA Kit (#ELK2007, Gentaur) according to the manufacturer’s instructions.

Techniques: Variant Assay, Immunofluorescence, Expressing, Isolation

Journal: Acta Neuropathologica Communications

Article Title: GJA1 (connexin43) is a key regulator of Alzheimer’s disease pathogenesis

doi: 10.1186/s40478-018-0642-x

Figure Lengend Snippet: Inflammatory cytokines downregulates Gja1, Apoe, and other network genes. a . Wildtype primary mouse astrocytes were treated with either IL-1β (10 ng/ml) or TNFα (10 ng/ml) or in combination for 7 days and levels of Cx43, Apoe expression were analyzed by immunoblot. Representative results from 4 ( Gja1 ) and 3 (Apoe) independent experiments are shown. b . Protein levels of Cx43 (left) and Apoe (right) were quantitatively analyzed for these technical replicates after normalization to Actin. ANOVA followed by Bonferroni post-hoc tests are indicated by asterisks. * p < 0.05, ** p < 0.01, *** p < 0.001. c . Quantitative gene expression analysis in wildtype astrocytes with treatments similar to a ) was performed to analyze the other key drivers of the GJA1 -centered network. Results representative of two independent experiments are shown. Statistical analysis was performed as in B, and only the comparison between control and IL-1β/TNFα is shown

Article Snippet: Mouse Apoe secreted in conditioned medium was quantitated using mouse Apoe ELISA kit (Mybiosource, MBS705227, San Diego, CA, USA) following manufacture’s instruction.

Techniques: Expressing, Western Blot, Gene Expression, Comparison, Control

Journal: Acta Neuropathologica Communications

Article Title: GJA1 (connexin43) is a key regulator of Alzheimer’s disease pathogenesis

doi: 10.1186/s40478-018-0642-x

Figure Lengend Snippet: GJA1 inhibitors downregulate Gja1 and Apoe, and other network genes. a . Wildtype primary mouse astrocytes were treated with 200 μM carbenoxolone (CBX, gap junction inhibitor) or Lanthanum (La3+, hemichannel inhibitor) for 3 days and levels of Cx43, Apoe were analyzed by immunoblot. b . Quantitative analysis of Cx43 and Apoe protein levels were normalized to Actin. ANOVA followed by Bonferroni post-hoc tests are indicated by asterisks. ** p < 0.01, *** p < 0.001. c . Quantitative gene expression analysis in wildtype astrocytes treated similar to A was performed to analyze GJA1 network drivers. Statistical analysis was performed as in b , and only the comparisons to control are shown. Results representative of two independent experiments are shown for all experiments

Article Snippet: Mouse Apoe secreted in conditioned medium was quantitated using mouse Apoe ELISA kit (Mybiosource, MBS705227, San Diego, CA, USA) following manufacture’s instruction.

Techniques: Western Blot, Gene Expression, Control

Journal: Acta Neuropathologica Communications

Article Title: GJA1 (connexin43) is a key regulator of Alzheimer’s disease pathogenesis

doi: 10.1186/s40478-018-0642-x

Figure Lengend Snippet: Gja1 deficiency causes reduction of Apoe production and promotes Aβ production in neuron/astrocyte cocultures. a . Quantitative PCR analysis of Apoe in wildtype (WT) and Gja1−/− astrocytes. b . Apoe, Cx43, and actin levels in wildtype and Gja1 −/− astrocytes treated with the indicated concentration of A β 1–42 oligomers for 48 h were determined by immunoblot analysis of cell lysates. c . Apoe secreted into the conditioned medium of astrocytes in B was quantitatively determined by specific ELISA in quadruplicate wells. d and e . Amyloidβ secreted to the conditioned medium of wildtype cortical neurons cocultured with either wildtype or Gja1−/− astrocytes was quantitatively determined by Aβ 40 and Aβ 42 specific ELISA

Article Snippet: Mouse Apoe secreted in conditioned medium was quantitated using mouse Apoe ELISA kit (Mybiosource, MBS705227, San Diego, CA, USA) following manufacture’s instruction.

Techniques: Real-time Polymerase Chain Reaction, Concentration Assay, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Acta Neuropathologica Communications

Article Title: GJA1 (connexin43) is a key regulator of Alzheimer’s disease pathogenesis

doi: 10.1186/s40478-018-0642-x

Figure Lengend Snippet: A working hypothesis of GJA1 dysregulation in AD. Progressive upregulation of GJA1 in the brains during LOAD pathogenesis (in response to amyloid accumulation) serves to enhance the coordinated gene network function to orchestrate neuroprotective response by astrocytes including Aβ production and clearance ( CST3, CLU, CTSB, ECE2, MEGF10, PSENEN and GSAP ), APOE production ( APOE , ABCA1, CAV1 and SPON1 ), and supporting neuronal activity ( DLG4, NLGN3, GRINA and GRIN2D ). However, the enhanced neuronal activity due to chronic and prolonged GJA1 upregulation triggers neuronal wear and eventual death

Article Snippet: Mouse Apoe secreted in conditioned medium was quantitated using mouse Apoe ELISA kit (Mybiosource, MBS705227, San Diego, CA, USA) following manufacture’s instruction.

Techniques: Activity Assay