Journal: bioRxiv

Article Title: Characterizing the SASP-Dependent Paracrine Spreading of Senescence Between Human Brain Cell Types

doi: 10.64898/2026.02.10.705129

Figure Lengend Snippet: A) Volcano plot of Nomic Bio Core Immune panel (268 cytokines) in DMSO and BrdU CM from astrocytes showing factors that are upregulated in BrdU CM treated (red) (log2fc > 1, p-value < 0.05) and downregulated in BrdU CM treated (blue) (log2fc < -1, p-value < 0.05). Black data points indicate factors that do not reach the threshold (−1 < log2fc < 1 | p-value > 0.05). B) Volcano plot of Nomic Bio Core Immune panel cytokines in DMSO and BrdU CM from endothelial cells showing factors that are upregulated in BrdU CM treated (red) (log2fc > 1, p-value < 0.05) and downregulated in BrdU CM treated (blue) (log2fc < -1, p-value < 0.05). Black data points indicate factors that do not reach the threshold (−1 < log2fc < 1 | p-value > 0.05). C) Volcano plot of Nomic Bio Core Immune panel cytokines in DMSO and BrdU CM from microglia showing factors that are upregulated in BrdU CM treated (red) (log2fc > 1, p-value < 0.05) and downregulated in BrdU CM treated (blue) (log2fc < -1, p-value < 0.05). Black data points indicate factors that do not reach the threshold (−1 < log2fc < 1 | p-value > 0.05). D) Volcano plot of Nomic Bio Core Immune panel cytokines in DMSO and BrdU CM from oligodendrocytes showing factors that are upregulated in BrdU CM treated (red) (log2fc > 1, p-value < 0.05) and downregulated in BrdU CM treated (blue) (log2fc < -1, p-value < 0.05). Black data points indicate factors that do not reach the threshold (−1 < log2fc < 1 | p-value > 0.05). E) Volcano plot of Nomic Bio Core Immune panel cytokines in DMSO and BrdU CM from neurons showing factors that are upregulated in BrdU CM treated (red) (log2fc > 1, p-value < 0.05) and downregulated in BrdU CM treated (blue) (log2fc < -1, p-value < 0.05). Black data points indicate factors that do not reach the threshold (−1 < log2fc < 1 | p-value > 0.05). F) Venn diagram showing the number of upregulated factors specifically in astrocyte BrdU CM and microglia BrdU CM (69) as well as the commonly upregulated factors . G) Z-score heatmap showing selected Core Immune panel (Nomic Bio) cytokine expression in all cell types (endothelial cells (pink), microglia (yellow), oligodendrocytes (green), astrocytes (purple), and neurons (blue)) based on nELISA analysis of CM from DMSO (grey) and BrdU (red) treated cell lines (n=3 replicates). H) Normalized expression of CCL2 based on human cytokine array in DMSO CM (grey) and BrdU CM (red) from astrocytes, endothelial cells, and microglia (n=4 replicates). Normalized expression of MIF based on human cytokine array in DMSO CM (grey) and BrdU CM (red) from astrocytes, endothelial cells, and microglia (n=4 replicates). I) Normalized nELISA signal of CCL2 in DMSO CM (grey) and BrdU CM (red) from astrocytes, endothelial cells, microglia, oligodendrocytes, and neurons (n=3 replicates). Normalized nELISA signal of MIF in DMSO CM (grey) and BrdU CM (red) from astrocytes, endothelial cells, microglia, oligodendrocytes, and neurons (n=3 replicates). Data was analyzed by two-way ANOVA with Tukey’s multiple comparisons test (H) and two-way ANOVA with Šídák’s multiple comparisons test (I). All graphs show mean with error bars depicting standard deviation (ns, p>0.05, *** p<0.001).

Article Snippet: Treatment with the CCL2 synthesis inhibitor Bindarit (MedChemExpress, Catalog No. HY-B0498) ( ) was used to specifically target CCL2 production in senescent cells.

Techniques: Expressing, Standard Deviation

Journal: bioRxiv

Article Title: Characterizing the SASP-Dependent Paracrine Spreading of Senescence Between Human Brain Cell Types

doi: 10.64898/2026.02.10.705129

Figure Lengend Snippet: A) Schematic depicting BulkSignalR pipeline which uses known ligand-receptor interactions and affected downstream pathways to analyze their activation based on our bulk RNAseq data from DMSO and BrdU treated human cell lines (created with BioRender). B) Venn diagram showing the number of receptors inferred from BulkSignalR to be activated across each of the five human cell types. Three receptors were identified in common between astrocytes (purple), endothelial cells (pink), and microglia (yellow) which were the cell types shown to be capable of receiving senescence signals and becoming SA β-gal positive: CXCR7, KREMEN2, and GIPR. Only CXCR7 was expressed in the cell types capable of entering secondary senescence (astrocytes, endothelial cells, microglia) ( , S3B). C) TPM expression values of CXCR7 , its ligand CXCL12 , and DPP4 which cleaves and inactivates CXCL12 in DMSO (grey) and BrdU (red) treated cell lines (n=3 replicates). D) Schematic of the four selected SASP inhibitors mechanisms of action: Bindarit is a CCL2 synthesis inhibitor which prevents p65 activation of the CCL2 gene at the promoter region, ISO-1 is a MIF antagonist, ACT-1004-1239 is a CXCR7 antagonist, and Sitagliptin inhibits DPP4 preventing its action of cleaving and inactivating CXCL12 (created with BioRender). Data was analyzed by two-way ANOVA with Tukey’s multiple comparisons test (C). All graphs show mean with error bars depicting standard deviation (ns, p>0.05, ** p<0.01, *** p<0.001).

Article Snippet: Treatment with the CCL2 synthesis inhibitor Bindarit (MedChemExpress, Catalog No. HY-B0498) ( ) was used to specifically target CCL2 production in senescent cells.

Techniques: Activation Assay, RNA sequencing, Expressing, Standard Deviation

Journal: bioRxiv

Article Title: Characterizing the SASP-Dependent Paracrine Spreading of Senescence Between Human Brain Cell Types

doi: 10.64898/2026.02.10.705129

Figure Lengend Snippet: A) TPM expression values of CCL2 receptors CCR1, CCR4, CCRL2, and ACKR1 in DMSO (grey) and BrdU (red) treated cell lines (n=3 replicates). B) TPM expression values of receptors KREMEN2 and GIPR in DMSO (grey) and BrdU (red) treated cell lines (n=3 replicates). Data was analyzed by two-way ANOVA with Tukey’s multiple comparisons test (A-B). All graphs show mean with error bars depicting standard deviation (ns, p>0.05, * p<0.05, ** p<0.01, *** p<0.001).

Article Snippet: Treatment with the CCL2 synthesis inhibitor Bindarit (MedChemExpress, Catalog No. HY-B0498) ( ) was used to specifically target CCL2 production in senescent cells.

Techniques: Expressing, Standard Deviation