human aortic smc  (ATCC)

Journal: bioRxiv

Article Title: Mechanical stretch regulates inflammatory signaling in human smooth muscle cells

doi: 10.1101/2025.10.29.685276

Figure Lengend Snippet: A. Outline of the experiment. Human aortic SMCs were exposed to static conditions (24 h), mechanical stretch (10%, 6h) or TNF stimulation (10 ng/mL, 24h). Uniform Manifold Approximation and Projection (UMAP) visualization of the phenotypic diversity of SMCs after static culture, stretch and TNF stimulation. Each dot represents a single cell. Nine cell clusters were identified in human aortic SMCs (0.3 resolution) as follows: C0 (fibromyocyte SMCs), C1 (inflammatory SMCs), C2 (proliferative M-phase), C3 (intermediate SMCs), C4 (proliferative S-phase), C5 (proliferative M-S-phase), C6 (contractile SMCs), C7 (apoptotic SMCs), C8 (endothelial-like SMCs). B. Dot plot showing the expression of three genes defining each cell cluster shown in A. Color scale indicates log2 expression, and the size represents the percentage of cells. C. Integration of publicly available human coronary and carotid atherosclerotic plaques scRNA-seq datasets – with human aortic SMCs from this study. This analysis revealed 7 mesenchymal cell clusters (pC0 to pC6). The p stands for plaque. Analysis of human aortic SMCs in culture are shown in the background (light grey cluster). D. Human aortic SMCs (C0 to C8) are shown in front and human atherosclerotic cell populations are shown in the background (light grey clusters).

Article Snippet: Human aortic SMCs (ATCC, PCS-100-012, male donor) were authenticated by ATCC and tested negative for mycoplasma contamination prior to experiments.

Techniques: Expressing

Journal: bioRxiv

Article Title: Selective reduction of KCNA4 in vulnerable glutamatergic-serotonin neurons of the dorsal raphe nucleus in Alzheimer’s Disease

doi: 10.1101/2025.10.17.683113

Figure Lengend Snippet: A: Graphical overview of the htau DREADDs experiment. Htau mice were intracranially administered hM3Dq (n = 14), hM4Di (n=14), or control virus (n = 9) at 2 months’ age. DREADDs agonist (CNO DHC) was administered via water bottle for 6 weeks. B , C : RNAscope assessment of Fos expression in virally transduced Tph2 + cells before CNO washout (acute) and after 7-day washout. Two data points were collected per mouse in ( C ). No statistical test was performed in ( C ), data were qualitatively compared. D,E : Novel objection recognition (NOR). D : Percent preference for novel object and E , discrimination index. 1w ANOVA with Tukey’s post hoc . F,G : Novelty-induced suppression of feeding (NSF). F : latency to feed and G , time spent in the corners of the arena during 10-minute assessment period. H–K : Elevated plus maze (EPM) task. H : Percent of total time spent in open arm, I , entries to open arm, J , total distance traveled, and K , average velocity during the task. L–R : Three-chamber social interaction test (SIT). L : Total social interaction time, M , number of social interactions, N , social interaction time as a percentage of time spent in the social chamber, and O , time spent in the center chamber. P–R Representative heat maps, normalized within trial. D–F,H–K : 1w ANOVA w/ Dunnett’s post hoc . G , 1w Welch’s ANOVA w/Dunnett’s post hoc . L-O : 1w ANOVA w/ Tukey’s post hoc .* p < 0.05, ** p < 0.01

Article Snippet: Coronal sections containing the DRN were mounted on histobond glass slides, blocked for 60’, and incubated in primary antibodies targeting Tph2 (goat, Everest EB11012, 1:375), VGLUT3 (mouse, Sigma-Aldrich SAB5200312, 1:500) phosphorylated human tau (pTau; AH36, rabbit, StressMarq SMC-601, 1:500) at 4°C overnight.

Techniques: Control, Virus, RNAscope, Expressing

Journal: bioRxiv

Article Title: Selective reduction of KCNA4 in vulnerable glutamatergic-serotonin neurons of the dorsal raphe nucleus in Alzheimer’s Disease

doi: 10.1101/2025.10.17.683113

Figure Lengend Snippet: A: Graphical overview of the htau DREADDs experiment. B–G: Confocal images of the DRN with immunolabeled Tph2 and pTau (AH36 antibody). Viral mCherry reporter ( top row ) was not amplified before imaging. One section per mouse that contained virally transduced cells was stained and imaged. H , left: Comparison of total virus coverage within the DRN expressed as percent immunoreactive area ( %IR ). Measurements were made for all mCherry+ area, Tph2 staining was used for anatomical alignment only. H, right: Assessment of pathology within the DRN, expressed as % IR for pTau immunolabeling within mCherry+ pixels of the DRN. I: Same as ( H ), but limited to the CM DRN. 1w ANOVA with Dunnett’s post hoc . ** p < 0.01

Article Snippet: Coronal sections containing the DRN were mounted on histobond glass slides, blocked for 60’, and incubated in primary antibodies targeting Tph2 (goat, Everest EB11012, 1:375), VGLUT3 (mouse, Sigma-Aldrich SAB5200312, 1:500) phosphorylated human tau (pTau; AH36, rabbit, StressMarq SMC-601, 1:500) at 4°C overnight.

Techniques: Immunolabeling, Amplification, Imaging, Staining, Comparison, Virus

Journal: bioRxiv

Article Title: Selective reduction of KCNA4 in vulnerable glutamatergic-serotonin neurons of the dorsal raphe nucleus in Alzheimer’s Disease

doi: 10.1101/2025.10.17.683113

Figure Lengend Snippet: A: Post hoc determination of 5HT/glut neurons using biocytin conjugation (green) in tandem with RNAscope targeting of Tph2 (red) and Slc17a8 (VGLUT3; magenta). B: Threshold current of 5HT/glut neurons, as determined using incremental current injections from -100–200 pA, P d = 1 s, interval 10 s. C: Input resistance measurements obtained at -60 mV holding potential. D–F: Current-clamp recordings obtained using the recording paradigm in ( B ), C57 vs htau 5HT/glut neurons, demonstrating event frequency ( D ), average inter-spike interval ( E ), and instantaneous frequency ( F ). 2w mixed model with RM. G–I: Same as D–F , but normalized within each recorded cell to compare current-dependent action-potential df/dt. B: Mann-Whitney test. C: Student’s t -test. D–I: 2w RM ANOVA w/Sidak’s post hoc . * p < 0.05, ** p < 0.01, **** p < 0.0001

Article Snippet: Coronal sections containing the DRN were mounted on histobond glass slides, blocked for 60’, and incubated in primary antibodies targeting Tph2 (goat, Everest EB11012, 1:375), VGLUT3 (mouse, Sigma-Aldrich SAB5200312, 1:500) phosphorylated human tau (pTau; AH36, rabbit, StressMarq SMC-601, 1:500) at 4°C overnight.

Techniques: Conjugation Assay, RNAscope, MANN-WHITNEY

Journal: bioRxiv

Article Title: Selective reduction of KCNA4 in vulnerable glutamatergic-serotonin neurons of the dorsal raphe nucleus in Alzheimer’s Disease

doi: 10.1101/2025.10.17.683113

Figure Lengend Snippet: A: Transcriptomically-distinct regions of the DRN as previously determined using Visium Spatial Gene Expression. B: Disease-relevant pathways affected by differentially-expressed genes (DEGs) in the centromedial DRN of htau mice as compared to wild-type mice. C: Hierarchical clustering of centromedial DEGs, top-50 delta only. Expression association is Z-scored for comparison across genes. D: Volcano plot of centromedial DRN DEGs, with ion channel genes annotated. E: Plot from ( D ), compared to the Agora database of human post-mortem Alzheimer’s DEGs. Genes that retain green/orange coloring were found to be differentially expressed in both Agora and our datasets, independent of their respective differential direction in human brain. F: Heatmap expression of Slc17a8 (VGLUT3) within the DRN from our Visium dataset. G: RNAscope labeling of Tph2 (green, left, marker for serotonin), Slc17a8 (magenta, center), and both (right) in the centromedial (CM) DRN.

Article Snippet: Coronal sections containing the DRN were mounted on histobond glass slides, blocked for 60’, and incubated in primary antibodies targeting Tph2 (goat, Everest EB11012, 1:375), VGLUT3 (mouse, Sigma-Aldrich SAB5200312, 1:500) phosphorylated human tau (pTau; AH36, rabbit, StressMarq SMC-601, 1:500) at 4°C overnight.

Techniques: Gene Expression, Expressing, Comparison, RNAscope, Labeling, Marker

Journal: bioRxiv

Article Title: Selective reduction of KCNA4 in vulnerable glutamatergic-serotonin neurons of the dorsal raphe nucleus in Alzheimer’s Disease

doi: 10.1101/2025.10.17.683113

Figure Lengend Snippet: A: Number of serotonergic cells ( Tph2 ), 5HT/glut cells ( Tph2 / Slc17a8 (VGLUT3)) and Kcna4 -(Kv1.4)-expressing 5HT/glut cells ( Tph2 / Slc17a8 / Kcna4 ) in the DRN, wild-type (C57) and htau mouse, one tissue section per animal. The percentage of triple-positive cells, respective of double-positive cells, was found to be reduced in htau mice. B: Confocal image demonstrating the expression of Tph2 , Slc17a8 , and Kcna4 with regards to the centromedial ( CM ) DRN; aq : aqueduct. C: Quantitation of Kcna4 puncta within all serotonergic cells ( Tph2 +, left) and 5HT/glut cells ( Tph2 +/ Slc17a8 +, right) of the DRN. D: Representative images of Kcna4 expression in 5HT/glut cells, C57 (left) and htau (right). E–H: same as A–D , but for Slc24a5 (NCKX5). I– L: same as A–D , but for Scn4b (Navβ4). A,E,I: 2w RM ANOVA w/Sidak’s post hoc . C,G: Student’s t -test. K: Welch’s t -test (left), Mann-Whitney test (right). * p < 0.05, ** p < 0.01, **** p < 0.0001

Article Snippet: Coronal sections containing the DRN were mounted on histobond glass slides, blocked for 60’, and incubated in primary antibodies targeting Tph2 (goat, Everest EB11012, 1:375), VGLUT3 (mouse, Sigma-Aldrich SAB5200312, 1:500) phosphorylated human tau (pTau; AH36, rabbit, StressMarq SMC-601, 1:500) at 4°C overnight.

Techniques: Expressing, Quantitation Assay, MANN-WHITNEY