Journal: Journal of Inflammation (London, England)

Article Title: Cinnamaldehyde inhibits the NLRP3 inflammasome by preserving mitochondrial integrity and augmenting autophagy in Shigella sonnei -infected macrophages

doi: 10.1186/s12950-024-00395-w

Figure Lengend Snippet: Role of autophagy in CA-mediated IL-1β inhibition in S. sonnei -infected macrophages. ( A-C ) J774A.1 macrophages were incubated with 40 µM CA for 4–20 h or 1 µM rapamycin for 4 h. The expression levels of LC3 ( A ), ATG5 ( B ) and p62 ( C ) in the cell lysates were analysed by Western blotting. ( D ) J774A.1 macrophages were primed for 4 h with LPS in the presence or absence of 1 mM 3-MA, followed by incubation with 40 µM CA for 0.5 h before S. sonnei infection for an additional 21 h. The detectable levels of IL-1β in the supernatants were analysed by ELISA. ( E ) Wild-type or LC3-knockdown J774A.1 macrophages were incubated with 40 µM CA for 20 h. The expression levels of LC3 in the cell lysates were analysed by Western blotting. ( F ) LPS-primed wild-type or LC3-knockdown J774A.1 macrophages were incubated with 40 µM CA for 0.5 h before S. sonnei infection for an additional 21 h. The detectable levels of IL-1β in the supernatants were analysed by ELISA. The Western blotting images are representative of separate experiments. The ELISA data are expressed as the means ± SD of three separate experiments. * p < 0.05 and *** p < 0.001 as indicated

Article Snippet: Mouse LC3 CRISPR/Cas9 knockout plasmids (sc-426,563 and sc-417,828-HDR) and antibodies against IL-18 (SC-6177), ASC (SC-22,514-R), cathepsin B (SC-365,558) and actin (SC-47,778) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Inhibition, Infection, Incubation, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Knockdown

Journal: bioRxiv

Article Title: Catestatin ameliorates tauopathy and amyloidogenesis via adrenergic inhibition

doi: 10.64898/2026.01.04.697519

Figure Lengend Snippet: (A) Experimental design showing intraperitoneal CST administration (0.5 µg/g BW) to PS19 mice from 5 to 9.5 months of age, followed by behavioral and pathological assessments. (B&C) CST preserves hippocampal structure in PS19 mice. ( B ) Representative Nissl-stained coronal sections and ( C ) volumetric analyses in nTg (n=8), Sal/PS19 (n=13), and CST/PS19 (n=11) reveal marked protection against hippocampal atrophy in CST-treated PS19 mice compared with saline-treated PS19 controls. (D–F) CST improves hippocampus-dependent working and spatial memory. In the Y-maze spontaneous alternation test (SAT), CST-treated PS19 mice exhibited ( D ) increased percent alternation and number of alterations (n=13). ( E ) In the two-trial Y-maze test, ( F ) CST-treated mice show greater novel-arm exploration than saline-treated PS19 mice (n=13). (G) Representative heatmaps of arm exploration illustrate superior spatial memory in CST-treated PS19 mice. (H–J) CST ameliorates motor deficits and restores innate behaviors. CST-treated PS19 mice show ( H ) improved grip strength (n=15), ( I ) significantly enhanced rotarod performance across training trials (nTg: n=19, Sal/PS19: n=18, CST/PS19: n=15), and ( J ) improved nest-building behavior compared with saline-treated PS19 animals (nTg: n=18, Sal/PS19: n=18, CST/PS19: n=15). *Data represent mean ± SEM. Significance levels: *p < 0.05, **p < 0.01, *** p < 0.001, ****p < 0.0001; NS, not significant

Article Snippet: Transgenic PS19 mice (Tau P301S; B6;C3-Tg(Prnp-MAPT P301S)PS19Vle/J) and 5xFAD mice (B6.Cg-Tg(APPSwFlLon,PSEN1 M146L*L286V)6799Vas/Mmjax) were purchased from The Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Staining, Saline

Journal: bioRxiv

Article Title: Catestatin ameliorates tauopathy and amyloidogenesis via adrenergic inhibition

doi: 10.64898/2026.01.04.697519

Figure Lengend Snippet: Immunoblot analysis of cortical lysates shows elevated phospho-Tau species (S202/T205, S396/404) in saline-treated PS19 mice, both of which are significantly reduced following CST treatment (nTg: n=2, Sal/PS19: n=5, CST/PS19: n=6). (D–F) CST exerts similar biochemical effects in the hippocampus, where CST administration markedly decreases pathological Tau phosphorylation at S202/T205 and S396/S404 in PS19 mice (nTg: n=2, Sal/PS19: n=6, CST/PS19: n=6). ( G&H) Hippocampal immunostaining with AT100 (recognizing pTau T212/S214) reveals substantial accumulation of pathological Tau in saline-treated PS19 mice; CST treatment significantly reduces the AT100-positive area, indicating reduced late-stage Tau pathology (nTg: n=3, Sal/PS19: n=5, CST/PS19: n=5). Scale bar = 200µm. ( I&J) CST attenuates misfolded Tau burden. MC1 immunostaining demonstrates robust conformational Tau pathology in saline-treated PS19 mice, whereas CST markedly diminishes MC1-positive aggregates throughout the hippocampus (nTg: n=3, Sal/PS19: n=5, CST/PS19: n=5). Scale bar = 200µm. ( K&L) CST suppresses Tau seeding competency. In HEK293 Tau-RD FRET biosensor assay, cortical lysates from saline-treated PS19 mice induce strong Tau aggregation. In contrast, lysates from CST-treated PS19 brains elicit significantly fewer GFP-positive aggregates, indicating reduced seeding activity (nTg: n=4, Sal/PS19: n=4, CST/PS19: n=4). Scale bar = 50µm. A.U.: Arbitrary Unit. Data represent mean ± SEM; significance indicated as *p < 0.05, **p < 0.01, *** p < 0.001, ****p < 0.0001; NS, not significant.

Article Snippet: Transgenic PS19 mice (Tau P301S; B6;C3-Tg(Prnp-MAPT P301S)PS19Vle/J) and 5xFAD mice (B6.Cg-Tg(APPSwFlLon,PSEN1 M146L*L286V)6799Vas/Mmjax) were purchased from The Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Western Blot, Saline, Phospho-proteomics, Immunostaining, Biosensor Assay, Activity Assay

Journal: bioRxiv

Article Title: Catestatin ameliorates tauopathy and amyloidogenesis via adrenergic inhibition

doi: 10.64898/2026.01.04.697519

Figure Lengend Snippet: UMAP visualization of snRNA-seq data. ( A ) Merged dataset showing annotated neuronal and glial clusters. ( B ) Non-transgenic (nTg) brains display normal cellular distributions. ( C ) Sal/PS19 mice show loss of CA1-ProS and SUB-ProS neurons and expansion of Astro and Micro-PVM clusters. ( D ) CST treatment partially restores CA1-ProS/SUB-ProS populations and reduces Astro and Micro-PVM clustering. (E&F) Number of Downregulated and Upregulated differentially expressed genes (DEG) in the reference group (top) vs comparison group (bottom) for the identified neuronal and glial clusters. ( G ) Enriched upregulated pathways with GO terms in CA1-ProS cluster of nTg and CST/PS19 compared to Sal/PS19. ( H ) Enriched upregulated pathways with GO terms in SUB-ProS cluster of nTg and CST/PS19 compared to Sal/PS19. (I&J) CD68 immunostaining depicts increased hippocampal microglial activation in Sal/PS19 mice, which is significantly reduced by CST (nTg: n=3, Sal/PS19: n=5, CST/PS19: n=5). Scale bars = 200µM. (K&L) GFAP staining reveals robust hippocampal astrogliosis in Sal/PS19 mice; CST markedly decreases GFAP⁺ area (nTg: n=3, Sal/PS19: n=5, CST/PS19: n=5). Scale bars = 200µM. (M&N) GFAP staining in the entorhinal cortex also shows elevated astrocytosis in Sal/PS19 mice, partially rescued by CST (nTg: n=3, Sal/PS19: n=5, CST/PS19: n=5). Scale bars = 200µM. Data are mean ± SEM; Significance indicated as *p < 0.05, **p < 0.01, *** p < 0.001, ****p < 0.0001; NS, not significant.

Article Snippet: Transgenic PS19 mice (Tau P301S; B6;C3-Tg(Prnp-MAPT P301S)PS19Vle/J) and 5xFAD mice (B6.Cg-Tg(APPSwFlLon,PSEN1 M146L*L286V)6799Vas/Mmjax) were purchased from The Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Transgenic Assay, Comparison, Immunostaining, Activation Assay, Staining

Journal: bioRxiv

Article Title: Catestatin ameliorates tauopathy and amyloidogenesis via adrenergic inhibition

doi: 10.64898/2026.01.04.697519

Figure Lengend Snippet: (A) Cortical epinephrine (EPI) levels are markedly elevated in Sal/PS19 compared with nTg controls, and CST treatment significantly lowers EPI concentrations (nTg: n=7, Sal/PS19: n=18, CST/PS19: n=15). (B) Cortical EPI levels are markedly elevated in Sal/5xFAD compared with nTg controls, and CST treatment significantly lowers EPI concentrations (nTg: n=7, Sal/5xFAD: n=6, CST/5xFAD: n=6). (C–E) CST suppresses pathological PKA signaling in the PS19 hippocampus. ( C ) Immunoblots show increased phospho-PKA substrates and elevated p-Tau (S202/T205) in Sal/PS19 mice; CST treatment reduces both, while total Tau and PKA-Cα remain unchanged. ( D&E ) Quantification confirms CST-mediated reduction of PKA activity and Tau phosphorylation (nTg: n=3, Sal/PS19: n=6, CST/PS19: n=5). (F) Schematic illustrating CST blockade ADR–driven PKA activation. In AAV-MAPT (P301S/K18)–transduced hippocampal slices, EPI strongly activates PKA signaling and enhances Tau phosphorylation, whereas CST antagonizes adrenergic/PKA hyperactivation and prevents formation of misfolded Tau species. (G–I) CST counteracts EPI-induced PKA activation and Tau pathology. ( G ) Immunoblot analysis shows that EPI markedly increases phospho-PKA substrates and p-Tau species (S202, S396/404), CST alone has minimal effect on PKA activity. Still, CST prevents EPI-induced PKA hyperactivation (PBS: n=3, CST: n=3, EPI: n=3, CST+EPI: n=3). ( H&I ) Quantification demonstrates CST’s ability to blunt adrenergic/PKA overactivation and its downstream Tau phosphorylation signatures. (J) Immunofluorescence staining (CA3, DG) reveals substantial accumulation of misfolded Tau in EPI-treated slices, which is significantly reduced by CST and nearly abolished by CST+EPI (PBS: n=3, CST: n=3, EPI: n=3, CST+EPI: n=3). Scale bar = 200µM. A.U.: Arbitrary Unit. Data shown as mean ± SEM; significance indicated as *p < 0.05, **p < 0.01, *** p < 0.001, ****p < 0.0001; NS, not significant.

Article Snippet: Transgenic PS19 mice (Tau P301S; B6;C3-Tg(Prnp-MAPT P301S)PS19Vle/J) and 5xFAD mice (B6.Cg-Tg(APPSwFlLon,PSEN1 M146L*L286V)6799Vas/Mmjax) were purchased from The Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Western Blot, Activity Assay, Phospho-proteomics, Activation Assay, Immunofluorescence, Staining