Journal: bioRxiv

Article Title: SPIN90 modulates the architecture of lamellipodial actin in an ARPC5L dependent fashion

doi: 10.64898/2025.12.01.691495

Figure Lengend Snippet: SPIN90 selectively activates ArpC5L containing Arp2/3 complexes (A) SPIN90 efficiently activates ArpC5L but not ArpC5 containing Arp2/3 complexes. Representative plots of pyrene-actin polymerization assays with 10 nM Arp2/3 complex, 100 nM SPIN90 and 2.5 µM actin (5 % pyrene labelled). It demonstrates that ArpC5L- but not ArpC5-containing Arp2/3 complexes activated by SPIN90 induce rapid actin polymerization, regardless of the ArpC1 isoform. The experiments were repeated independently three times and yielded similar results. (B) Representative TIRF images demonstrate that ArpC5L- but not ArpC5-containing complex activated by SPIN90 efficiently generate actin filaments. Scale bar = 20 µm A mixture of 20 nM Arp2/3 complex, 200 nM SPIN90 and 0.5 µM actin (15 % Alexa 488 labelled) was observed overtime. The graph shows quantitative analysis of filament density over time from three independent technical replicates. The points indicate the mean density, and the error bars represent the standard deviation.

Article Snippet: Other antibodies are including: ARPC2/p34-Arc polyclonal (Millipore, 07-227), ARPC5 monoclonal (Synaptic Systems, 305011), ArpC5L (Proteintech, 22025-a-AP), β-actin (Abcam, ab179467), anti-mouse IgG-IRDye® 800CW (LI-COR Biosciences, 926-32210) and anti-rabbit IgG-IRDye® 680RD (LI-COR Biosciences, 926-68071).

Techniques: Standard Deviation

Journal: bioRxiv

Article Title: SPIN90 modulates the architecture of lamellipodial actin in an ARPC5L dependent fashion

doi: 10.64898/2025.12.01.691495

Figure Lengend Snippet: SPIN90 has a higher affinity for Arp2/3 complexes with ArpC5L than ArpC5 (A) Surface representation of the Arp2/3 complex from the SPIN90-Arp2/3 complex structure (PDB: 6DEC). Residues in Arp2/3 complex subunits contacting the two SPIN90 molecules are shown in darker colours. (B) Immunoblots using an ArpC2 antibody show the amount of Arp2/3 complex containing ArpC1B pulled down with GST–SPIN90-Cter. Both blots were run on the same gel and developed on the same membrane to allow a direct comparison of signal intensity. (C) The graph shows the quantification of average normalized ArpC2 intensity from three independent replicates, with error bars resenting standard deviation. The data was fit with Hill equation.

Article Snippet: Other antibodies are including: ARPC2/p34-Arc polyclonal (Millipore, 07-227), ARPC5 monoclonal (Synaptic Systems, 305011), ArpC5L (Proteintech, 22025-a-AP), β-actin (Abcam, ab179467), anti-mouse IgG-IRDye® 800CW (LI-COR Biosciences, 926-32210) and anti-rabbit IgG-IRDye® 680RD (LI-COR Biosciences, 926-68071).

Techniques: Western Blot, Membrane, Comparison, Standard Deviation

Journal: bioRxiv

Article Title: SPIN90 modulates the architecture of lamellipodial actin in an ARPC5L dependent fashion

doi: 10.64898/2025.12.01.691495

Figure Lengend Snippet: SPIN90 localizes at the leading edge and integrates into lamellipodial actin networks. (A) Quantitative immunoblot analysis of endogenous levels of ArpC5 isoforms in B16 cells using recombinant His-tagged ArpC5 and His-tagged ArpC5L as standards. The average concentrations and standard deviations from three independent replicates are shown in the table on the right. (B) Schematic shows the two Ruby-tagged SPIN90 constructs expressed in cells. Representative live images show B16 cells overexpressing GFP (volume marker) together with Ruby–SPIN90 (upper) or with Ruby-SPIN90 Nter (lower). Scale bars = 5 µm. Signal intensity (right) is quantified by normalizing the intensity at the leading edge (solid line) compared to that 2□µm from the leading edge (dashed line). Each pair of points represents measurements from the same cell with the different colours representing data from three independent replicates. A two-tailed paired t-test is used to estimate the p-value.

Article Snippet: Other antibodies are including: ARPC2/p34-Arc polyclonal (Millipore, 07-227), ARPC5 monoclonal (Synaptic Systems, 305011), ArpC5L (Proteintech, 22025-a-AP), β-actin (Abcam, ab179467), anti-mouse IgG-IRDye® 800CW (LI-COR Biosciences, 926-32210) and anti-rabbit IgG-IRDye® 680RD (LI-COR Biosciences, 926-68071).

Techniques: Western Blot, Recombinant, Construct, Marker, Two Tailed Test

Journal: bioRxiv

Article Title: SPIN90 modulates the architecture of lamellipodial actin in an ARPC5L dependent fashion

doi: 10.64898/2025.12.01.691495

Figure Lengend Snippet: SPIN90 enhances the recruitment of ArpC5L but not ArpC5 to the leading edge Representative images of live cells expressing endogenous NeonGreen–ArpC5L (A) or ArpC5-NeonGreen (B) after treatment with the indicated siRNAs. Allstar represents siRNA control. The graphs show quantification of the ArpC5L or ArpC5 signal intensity at the leading edge in the different conditions. Each point represents an individual cell, with different colours indicating 3 independent experimental replicates. For each repeat, 8-15 cells were imaged and quantified. The mean of each experiment is shown as a circle, and error bars are standard deviation. A two-tailed paired t-test was used to calculate the p-value. Scale bars = 5 µm

Article Snippet: Other antibodies are including: ARPC2/p34-Arc polyclonal (Millipore, 07-227), ARPC5 monoclonal (Synaptic Systems, 305011), ArpC5L (Proteintech, 22025-a-AP), β-actin (Abcam, ab179467), anti-mouse IgG-IRDye® 800CW (LI-COR Biosciences, 926-32210) and anti-rabbit IgG-IRDye® 680RD (LI-COR Biosciences, 926-68071).

Techniques: Expressing, Control, Standard Deviation, Two Tailed Test

Journal: bioRxiv

Article Title: SPIN90 modulates the architecture of lamellipodial actin in an ARPC5L dependent fashion

doi: 10.64898/2025.12.01.691495

Figure Lengend Snippet: The organisation of actin filaments at the leading edge of B16 cells. (A) Schematic representation of ρ (rho) and ψ (psi) in the organization of the actin network. (B) Representative images of an Alexa-488 phalloidin stained B16 cell for each cell line as indicated. The coloured region in the lamellipodium is the result of polarimetry analysis within a 0.65 µm (10-pixel) region from the leading edge to determine the mean actin filament orientation in each pixel. The mean filament orientation per pixel is represented with a stick whose orientation corresponds to filament orientation (angle rho, ρ) and whose color depicts filament orientation with respect to the leading edge (angle rho c , ρ c ) within the 0–180° range according to the colour bar. The stick maps of a representative ROI (labelled with white rectangles) are zoomed in and shown. (C) The polar histogram shows the statistical distribution of actin filament angles (ρ c ) per pixel within the 0–90° range relative to the leading edge of all the cells, with 38 to 55 cells analysed per condition across three independent repeats. The average < ρ c > and standard deviation for each cell line are indicated. The colours correspond to the colour bar shown in panel A. (D) Violin plot showing the statistical distribution of actin filament angles (ρ c ) per pixel within the 0–90° range, as shown in panel B. A two-tailed Mann–Whitney test was used to calculate the p-value between individual cells under two conditions. The medians and interquartile ranges are shown as black lines, while the means and standard deviations are shown as blue dots with error bars. (E) The statistical results of the average lamellipodial actin filament angle < ρ c > relative to the leading edge within the 0–90° range in individual wildtype, ArpC5, ArpC5Lor SPIN90 knockout cells. Each point represents the mean angle of actin filaments relative to the leading edge in a single cell from three independent experiments, the latter shown in different colours. Mean ± SD for each experimental condition is indicated. A two-tailed Welch’s t-test is used to estimate the p-value between the means under two conditions. (F) The statistical results of the standard deviation (SD) of actin filament angles relative to the leading edge within the 0–90° range in individual wildtype, ArpC5, ArpC5L, or SPIN90 knockout cells. Each point represents the SD of actin filaments relative to the leading edge in a single cell from three independent experiments, the latter shown in different colours. Mean ± SD for each experimental condition is indicated. A two-tailed Mann Whitney test is used to estimate the p-value between individual cells under two conditions. (G) Representative images of Alexa-488 phalloidin stained B16 cell (right) overexpressing Ruby-SPIN90 (left). The zoomed-in image shows the mean actin filament orientation per pixel within the ROI (white rectangle) as in panel A. (H) The polar histogram shows the statistical distribution of actin filament angles (ρ c ) per pixel within the 0–90° range relative to the leading edge in all cells overexpressing either Ruby-SPIN90 (61 cells) or Ruby-SPIN90 Nter (59 cells), across three independent experimental repeats. The average < ρ c > and standard deviation for each experimental condition are indicated. The colours correspond to the colour bar shown in panel A.

Article Snippet: Other antibodies are including: ARPC2/p34-Arc polyclonal (Millipore, 07-227), ARPC5 monoclonal (Synaptic Systems, 305011), ArpC5L (Proteintech, 22025-a-AP), β-actin (Abcam, ab179467), anti-mouse IgG-IRDye® 800CW (LI-COR Biosciences, 926-32210) and anti-rabbit IgG-IRDye® 680RD (LI-COR Biosciences, 926-68071).

Techniques: Staining, Standard Deviation, Two Tailed Test, MANN-WHITNEY, Knock-Out

Journal: bioRxiv

Article Title: SPIN90 modulates the architecture of lamellipodial actin in an ARPC5L dependent fashion

doi: 10.64898/2025.12.01.691495

Figure Lengend Snippet: SPIN90 impacts the lamellipodia protrusion and cell motility. (A) Representative image stills of WT, ArpC5, ArpC5Land SPIN90 knockout B16 cells expressing endogenously tagged GFP β-actin. (B) The quantification of lamellipodia width, lamellipodial actin density. Each point represents an individual cell, with different colours indicating independent experimental replicates. The mean of each experiment is shown as a circle, and error bars are standard deviation. A two-tailed paired t-test was used to estimate the p-value between the means under two conditions. (C) Representative image still (upper) from a FRAP assay on a B16 cell with endogenously GFP tagged β–actin. The kymograph (lower) shows the cell protrusion (indicated by straight line) and rearward flow (indicated by dashed line), made visible by the FRAP region. (D) The quantification of lamellipodial protrusion rate, rearward flow and protrusion efficiency. Each point represents an individual cell, with different colours indicating independent experimental replicates. The mean of each experiment is shown as a circle, and error bars are standard deviation. A two-tailed paired t-test was used to estimate the p-value between the means under two conditions. (E) Quantification of B16 cell random migration speed. Each point represents an individual cell, with different colours indicating 3 independent replicates. The mean of each experiment is shown as a circle, and error bars represent standard deviation. A two-tailed paired t-test was used to estimate the p-value between the means under two conditions. (F) Representative images (left panels) of the leading edge of B16 cells overexpressing either Ruby, or Ruby-SPIN90. The right panel shows the protrusion rate of these cells. Each point represents an individual cell, with different colours indicating independent experimental replicates. The mean of each experiment is shown as a circle, and error bars are standard deviation. A two-tailed paired t-test was used to estimate the p-value between the means under two conditions.

Article Snippet: Other antibodies are including: ARPC2/p34-Arc polyclonal (Millipore, 07-227), ARPC5 monoclonal (Synaptic Systems, 305011), ArpC5L (Proteintech, 22025-a-AP), β-actin (Abcam, ab179467), anti-mouse IgG-IRDye® 800CW (LI-COR Biosciences, 926-32210) and anti-rabbit IgG-IRDye® 680RD (LI-COR Biosciences, 926-68071).

Techniques: Knock-Out, Expressing, Standard Deviation, Two Tailed Test, FRAP Assay, Migration

Journal: bioRxiv

Article Title: SPIN90 modulates the architecture of lamellipodial actin in an ARPC5L dependent fashion

doi: 10.64898/2025.12.01.691495

Figure Lengend Snippet: Schematic summary (A) In migrating cells, SPIN90 recruits the ArpC5L-, but not the ArpC5-containing complex, to the lamellipodial leading edge . Meanwhile, both ArpC5L- and ArpC5-containing complexes generate branches at the lamellipodia . Thus, SPIN90 and Arp2/3 iso-complexes integrate into the lamellipodial actin network, contributing to the formation of a complex actin architecture that ensures efficient protrusion . (B) ArpC5 KO cells (left panel), the ArpC5L-containing complex participates in the formation of both linear and branched actin filaments. Consequently, the distribution of filament orientations in their lamellipodia is much fig than that of the wild type. In contrast, in ArpC5L KO cells (right panel), the ArpC5-containing complex generates only branched actin filaments, resulting in a narrower distribution of filament orientations.

Article Snippet: Other antibodies are including: ARPC2/p34-Arc polyclonal (Millipore, 07-227), ARPC5 monoclonal (Synaptic Systems, 305011), ArpC5L (Proteintech, 22025-a-AP), β-actin (Abcam, ab179467), anti-mouse IgG-IRDye® 800CW (LI-COR Biosciences, 926-32210) and anti-rabbit IgG-IRDye® 680RD (LI-COR Biosciences, 926-68071).

Techniques:

Journal: bioRxiv

Article Title: Heme drives cardiac endothelial senescence in sepsis via STING activation

doi: 10.1101/2025.06.18.660159

Figure Lengend Snippet: ( A ) Gene set enrichment analysis (GSEA) of RNA-seq data from hearts of sham and septic mice, highlighting enrichment of senescence-related genes. ( B ) Heat map showing differential expression of selected senescence-associated genes. ( C ) Expression levels of representative senescence-related genes (Cdkn1a/p21, Cxcl2, Mmp9, Thbs1, Cxcl1) based on RNA-seq analysis of hearts from sham and septic mice. ( D ) Western blot analysis of senescence markers p16, p21, and acetylated p53 in sham and septic heart tissues (n=6 mice/group). ( E ) qRT-PCR analysis of senescence markers in sham and septic heart tissues (n=6-8 mice/group). ( F, G ) Western blot analysis and quantification of senescence markers p16, p21, and acetylated p53 in heart tissues from sham mice, septic non-survivors (body temperature < 30L°C), and septic survivors (> 30L°C) at 24 hours post-CLP (n=6 mice/group). ( H) qRT-PCR analysis of senescence associated genes (Cxcl9, Il-1A, Serpine1, Thbs1) in heart tissues from sham mice, septic survivors, and septic non-survivors. All data are presented as mean ± SD. Statistical significance was determined using the unpaired two-tailed Student’s t-test: *PL

Article Snippet: Anti-p16 (Cat: A8571), Anti-AP53 (Cat: A19836), Anti-Hemopexin (HPX) (Cat: A5603), Anti-DDDDK-Tag (Cat: AE005) were purchased from Abclonal.

Techniques: RNA Sequencing, Quantitative Proteomics, Expressing, Western Blot, Quantitative RT-PCR, Two Tailed Test

Journal: bioRxiv

Article Title: Heme drives cardiac endothelial senescence in sepsis via STING activation

doi: 10.1101/2025.06.18.660159

Figure Lengend Snippet: Cardiac endothelial cells are the predominant senescent population during sepsis . ( A-D ) Co-immunostaining of senescence markers p21 and p16 with the endothelial cell marker CD31, and their quantification in heart sections from sham and septic mice (n=6 mice/group). Scale bars, 50Lμm. ( E-F ) Co-immunostaining of the cellular proliferation marker Ki67 with the endothelial marker CD31 in heart tissues from sham controls and septic mice at 24 hours post-CLP, along with corresponding quantification. (n=8/group). Scale bars, 50Lμm. ( G ) β-gal staining of HUVECs treated with plasma (1:200) from sham or septic (CLP 24 h) mice for 24 hours (n=6/group). Scale bars, 100Lμm. ( H ) Quantification of β-gal staining in HUVECs, as shown in ( G ). ( I ) BrdU incorporation assay in HUVECs from the indicated treatment groups (n=6/group). Scale bars, 100Lμm. ( J) Quantification of BrdU incorporation in HUVECs, as shown in ( H ). All data are presented as mean ± SD. Statistical analysis was performed using the unpaired two-tailed Student’s t-test: *PL

Article Snippet: Anti-p16 (Cat: A8571), Anti-AP53 (Cat: A19836), Anti-Hemopexin (HPX) (Cat: A5603), Anti-DDDDK-Tag (Cat: AE005) were purchased from Abclonal.

Techniques: Immunostaining, Marker, Staining, Clinical Proteomics, BrdU Incorporation Assay, Two Tailed Test

Journal: bioRxiv

Article Title: Heme drives cardiac endothelial senescence in sepsis via STING activation

doi: 10.1101/2025.06.18.660159

Figure Lengend Snippet: Cardiac endothelial senescence impairs proliferation in sepsis . ( A-B ) Immunofluorescent co-staining and quantification of the senescence marker p21 (red) and the endothelial marker CD31 (green) in heart sections from the indicated groups (n=6 mice/group). Scale bars, 50Lμm. ( C-D ) Immunofluorescent co-staining and quantification of the senescence marker p16 (red) and the endothelial marker CD31 (green) in heart sections from the indicated groups (n=6 mice/group). Scale bars, 50Lμm. ( E, F ) Immunofluorescent co-staining and quantification of the proliferation marker Ki67 (red) and the endothelial marker CD31 (green) in heart sections from sham, septic survivors, and septic non-survivors (n=6-8 mice/group). Scale bars, 50Lμm. All data are presented as mean ± SD. Statistical analysis was performed using the unpaired two-tailed Student’s t-test: *PL

Article Snippet: Anti-p16 (Cat: A8571), Anti-AP53 (Cat: A19836), Anti-Hemopexin (HPX) (Cat: A5603), Anti-DDDDK-Tag (Cat: AE005) were purchased from Abclonal.

Techniques: Staining, Marker, Two Tailed Test

Journal: bioRxiv

Article Title: Heme drives cardiac endothelial senescence in sepsis via STING activation

doi: 10.1101/2025.06.18.660159

Figure Lengend Snippet: Elevated heme levels correlate with cardiac endothelial senescence and dysfunction in sepsis . ( A ) Plasma heme levels in sham mice, septic non-survivors (body temperature < 30L°C), and septic survivors (> 30L°C) at 24 hours post-CLP (n=6–8 mice/group). ( B, C ) Mice with higher circulating heme levels exhibited increased cardiac endothelial senescence, as indicated by elevated expression of senescence markers p21 and p16. ( D, E ) Co-immunostaining of p21 with endothelial cells marker CD31 and quantification in heart tissues from sham and septic mice with or without heme treatment (n=6 mice/group). Scale bars, 50Lμm. ( F–H ) Echocardiographic assessment of ejection fraction (EF) and fractional shortening (FS) in sham and septic mice treated with or without heme (n=6 mice/group). ( I, J ) Western blot analysis and quantification of senescence markers p16, p21, and acetylated p53 in heart tissues from sham and septic mice with or without heme treatment (n=6 mice/group). All data are presented as mean ± SD. Statistical analysis was performed using the unpaired two-tailed Student’s t-test: *PL

Article Snippet: Anti-p16 (Cat: A8571), Anti-AP53 (Cat: A19836), Anti-Hemopexin (HPX) (Cat: A5603), Anti-DDDDK-Tag (Cat: AE005) were purchased from Abclonal.

Techniques: Clinical Proteomics, Expressing, Immunostaining, Marker, Western Blot, Two Tailed Test

Journal: bioRxiv

Article Title: Heme drives cardiac endothelial senescence in sepsis via STING activation

doi: 10.1101/2025.06.18.660159

Figure Lengend Snippet: Heme administration enhances cardiac endothelial senescence and impairs endothelial proliferation . ( A, B ) Immunofluorescent co-staining and quantification of the senescence marker p16 (red) and the endothelial marker CD31 (green) in heart sections from the indicated groups (n=6 mice/group). Scale bars, 50Lμm. ( C, D ) Immunofluorescent co-staining and quantification of the proliferation marker Ki67 (red) and the endothelial marker CD31 (green) in heart sections from the indicated groups (n=6 mice/group). Scale bars, 50Lμm. All data are presented as mean ± SD. Statistical analysis was performed using the unpaired two-tailed Student’s t-test: *PL

Article Snippet: Anti-p16 (Cat: A8571), Anti-AP53 (Cat: A19836), Anti-Hemopexin (HPX) (Cat: A5603), Anti-DDDDK-Tag (Cat: AE005) were purchased from Abclonal.

Techniques: Staining, Marker, Two Tailed Test

Journal: bioRxiv

Article Title: Heme drives cardiac endothelial senescence in sepsis via STING activation

doi: 10.1101/2025.06.18.660159

Figure Lengend Snippet: Heme exacerbates bacterial induced endothelial senescence . ( A, B ) β-galactosidase staining and quantification of HUVECs treated with heme (10LµM), heat-killed E. coli (MOI:10), or both for 24 hours (n=6/group). Scale bars, 100Lμm. ( C, D ) Western blot analysis and quantification of senescence markers p16, p21, and acetylated p53 in the indicated HUVEC groups (n=4/group). ( E, F ) Immunofluorescent staining and quantification of p21 in the indicated HUVEC groups (n=6/group). Scale bars, 50Lμm. ( G, H ) BrdU incorporation assay and quantification in the indicated HUVEC groups (n=6/group). Scale bars, 100Lμm. ( I ) qRT-PCR analysis of senescence-associated genes in the indicated HUVEC groups (n=6/group). All data are presented as mean ± SD. Statistical analysis was performed using the unpaired two-tailed Student’s t-test: *PL

Article Snippet: Anti-p16 (Cat: A8571), Anti-AP53 (Cat: A19836), Anti-Hemopexin (HPX) (Cat: A5603), Anti-DDDDK-Tag (Cat: AE005) were purchased from Abclonal.

Techniques: Staining, Western Blot, BrdU Incorporation Assay, Quantitative RT-PCR, Two Tailed Test

Journal: bioRxiv

Article Title: Heme drives cardiac endothelial senescence in sepsis via STING activation

doi: 10.1101/2025.06.18.660159

Figure Lengend Snippet: STING activation contributes to sepsis induced cardiac endothelial senescence . ( A, B ) Co-immunofluorescent staining and quantification of the senescence marker p21 with the endothelial cell marker CD31 in cardiac tissues from sham and septic mice treated with a STING inhibitor or vehicle control at 24 hours post-CLP (n=6 mice/group). Scale bars, 50Lμm. ( C ) Representative echocardiographic images assessing ejection fraction (EF) and fractional shortening (FS) at 24 hours, 7 days, and 14 days post-CLP in mice treated with vehicle or STING inhibitor. ( D ) Western blot analysis of p-STING/STING, p-TBK1/TBK1, and p-IRF3/IRF3 in HUVECs treated with heme + heat-killed E. coli , with or without the STING inhibitor C-176 (2.5LμM) (n=6/group). ( E, F ) Echocardiographic assessment of EF and FS in septic mice treated with STING inhibitor or vehicle control at 24 hours, 7 days, and 14 days post-CLP, as shown in (C) (n=6 mice/group). ( G ) Quantification of Western blot analysis of p-STING/STING, p-TBK1/TBK1, and p-IRF3/IRF3 in the indicated HUVEC groups, as shown in ( D ) (n=4/group). ( H, I ) Western blot analysis and quantification of senescence markers p16, p21, and acetylated p53 in the indicated HUVEC groups (n=6/group). ( J, K ) β-galactosidase (β-gal) staining and quantification of HUVECs treated with combined heme and E. coli with or without the STING inhibitor C-176 (n=6/group). Scale bars, 100Lμm. All data are presented as mean ± SD. Statistical analysis was performed using the unpaired two-tailed Student’s t-test: *PL

Article Snippet: Anti-p16 (Cat: A8571), Anti-AP53 (Cat: A19836), Anti-Hemopexin (HPX) (Cat: A5603), Anti-DDDDK-Tag (Cat: AE005) were purchased from Abclonal.

Techniques: Activation Assay, Staining, Marker, Control, Western Blot, Two Tailed Test

Journal: bioRxiv

Article Title: Heme drives cardiac endothelial senescence in sepsis via STING activation

doi: 10.1101/2025.06.18.660159

Figure Lengend Snippet: STING inhibition attenuates sepsis-induced cardiac endothelial senescence . ( A, B ) Immunofluorescent co-staining and quantification of the senescence marker p16 with the endothelial marker CD31 in heart tissues from sham and septic mice treated with a STING inhibitor or vehicle at 24 hours post-CLP (n=5-6 mice/group). Scale bars, 50Lμm. ( C–E ) Immunofluorescent co-staining and quantification of senescence markers p16 or p21 with the endothelial marker CD31 in heart tissues from sham and septic mice treated with a STING inhibitor or vehicle at 14 days post-CLP (n=4-6 mice/group). Scale bars, 50Lμm. All data are presented as mean ± SD. Statistical analysis was performed using the unpaired two-tailed Student’s t-test: **PL

Article Snippet: Anti-p16 (Cat: A8571), Anti-AP53 (Cat: A19836), Anti-Hemopexin (HPX) (Cat: A5603), Anti-DDDDK-Tag (Cat: AE005) were purchased from Abclonal.

Techniques: Inhibition, Staining, Marker, Two Tailed Test

Journal: bioRxiv

Article Title: Heme drives cardiac endothelial senescence in sepsis via STING activation

doi: 10.1101/2025.06.18.660159

Figure Lengend Snippet: Increased hemopexin expression attenuates sepsis-induced cardiac endothelial senescence . ( A–D ) Immunostaining and quantification of the senescence markers p21 or p16 with endothelial cell markers in sham and septic heart tissues from AAV-HPX and AAV-Con mice at 24 hours post-CLP (n=6-7/group). Scale bars, 50Lμm. All data are presented as mean ± SD. Statistical analysis was performed using the unpaired two-tailed Student’s t-test: **PL

Article Snippet: Anti-p16 (Cat: A8571), Anti-AP53 (Cat: A19836), Anti-Hemopexin (HPX) (Cat: A5603), Anti-DDDDK-Tag (Cat: AE005) were purchased from Abclonal.

Techniques: Expressing, Immunostaining, Two Tailed Test

Journal: bioRxiv

Article Title: Heme drives cardiac endothelial senescence in sepsis via STING activation

doi: 10.1101/2025.06.18.660159

Figure Lengend Snippet: Increased hemopexin expression attenuates sepsis-induced cardiac endothelial senescence and enhances endothelial proliferative capacity . ( A, B ) Co-immunostaining and quantification of the cellular proliferation marker Ki67 with the endothelial marker CD31 in sham and septic heart tissues from AAV–HPX and AAV–Con mice at 24 hours post-sepsis. Scale bars, 50Lμm. ( C, D ) Co-immunostaining and quantification of the senescence marker p16 with the endothelial marker CD31 in heart tissues from AAV–HPX and AAV–Con mice at 14 days post-sepsis. Scale bars, 50Lμm. All data are presented as mean ± SD. Statistical analysis was performed using the unpaired two-tailed Student’s t-test: *PL

Article Snippet: Anti-p16 (Cat: A8571), Anti-AP53 (Cat: A19836), Anti-Hemopexin (HPX) (Cat: A5603), Anti-DDDDK-Tag (Cat: AE005) were purchased from Abclonal.

Techniques: Expressing, Immunostaining, Marker, Two Tailed Test