Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A ) Schematic diagram showing the strategy for the drug screen. B ) THP-1 ASC-GFP monocytic cells without any treatment. C ) THP-1 ASC-GFP monocytic cells treated with LPS (1μg/ml) for 2 hours. D ) THP-1 ASC-GFP monocytic cells treated with LPS (1μg/ml) for 4 hours and Nigericin (10 μM) for 1 hour.

Article Snippet: THP-1-ASC-GFP cells (Invivogen #thp-ascgfp) were maintained in RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum, 100 μg/ml NormocinTM,1% Pen/Strep.

Techniques:

Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A ) Vehicle-treated or drug-treated, 20 μM, 16h ( B-P ) THP-1 ASC-GFP monocytic cells were treated with LPS (1μg/ml) for 4 hours. THP-1 ASC-GFP cells were pretreated with selected drugs for 16h, followed by washing with PBS and incubation with LPS (1μg/ml) for 2 hours.

Article Snippet: THP-1-ASC-GFP cells (Invivogen #thp-ascgfp) were maintained in RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum, 100 μg/ml NormocinTM,1% Pen/Strep.

Techniques: Incubation

Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A ) THP-1 ASC-GFP cells pretreated with vehicle or selected drugs for 16h, followed by washing with PBS and incubation with LPS (1μg/ml) for 2 hours, followed by incubation with Nigericin (10 μM) for 1 hour. ASC-puncta were visualized using fluorescent microscopy, with DAPI used to stain nuclei. B ) RAW-ASC cells pretreated with vehicle or selected drugs for 16h, followed by washing with PBS and incubation with LPS (1μg/ml) for 2 hours, followed by incubation with Nigericin (5 μM) for 1 hour. Cells were fixed and permeabilized and stained with anti-ASC antibody and Alexa-labeled secondary antibody. ASC-puncta were visualized using fluorescent microscopy, with DAPI used to stain nuclei.

Article Snippet: THP-1-ASC-GFP cells (Invivogen #thp-ascgfp) were maintained in RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum, 100 μg/ml NormocinTM,1% Pen/Strep.

Techniques: Incubation, Microscopy, Staining, Labeling

Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A, B ) Western blot analysis of basal or LPS-induced expression of various inflammasome components in THP-1 ASC-GFP cells pretreated with vehicle or selected drugs for 16h. C, D ) Western blot analysis of basal or LPS-induced expression of various inflammasome components in RAW- ASC cells pretreated with vehicle or selected drugs for 16h. E ) Age-matched (10-week-old) male WTC57BL6J mice fed with chow diet were i.p injected with saline or saquinavir. After 2 hours, mice were primed for inflammasome assembly by an I.P. injection of LPS (5μg/mouse). After 4h of LPS injection, the NLRP3 inflammasome assembly was induced by I.P. injection of ATP (0.5 mL of 30 mM, pH 7.0). The peritoneal cavity was lavaged with 5 mL sterile PBS, and IL-1β levels in peritoneal lavage were determined by ELISA (N = 6, mean ± SD for all groups, ∗∗p < 0.01 by two-tailed t-test). Mouse plasma was used in multiplex analysis to determine levels of F ) TNF-α, G ) IL-1β, H ) IL-17A, I ) IL-33, and J ) IL-15 (N=3, mean ± SD for all groups, ∗p < 0.05 by two-tailed t-test).

Article Snippet: THP-1-ASC-GFP cells (Invivogen #thp-ascgfp) were maintained in RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum, 100 μg/ml NormocinTM,1% Pen/Strep.

Techniques: Western Blot, Expressing, Injection, Saline, Sterility, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Clinical Proteomics, Multiplex Assay

Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A ) THP-1 ASC-GFP macrophages pretreated with vehicle or selected drugs for 16h, followed by washing with PBS and incubation with LPS (1μg/ml) for 2 hours, followed by incubation with Mitosox Red dye and fluorescent microscopy. DAPI staining is used to visualize nuclei.

Article Snippet: THP-1-ASC-GFP cells (Invivogen #thp-ascgfp) were maintained in RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum, 100 μg/ml NormocinTM,1% Pen/Strep.

Techniques: Incubation, Microscopy, Staining

Journal: bioRxiv

Article Title: Recessive POPDC1 Truncation Causes Lethal Short-QT Pattern Arrhythmogenic Cardiomyopathy with Multi-Ion Channel Remodeling and Ankyrin-G Scaffold Disruption

doi: 10.64898/2026.03.07.710328

Figure Lengend Snippet: (A) Volcano plot depiction of the differential proteome from ventricular tissue lysates of Popdc1 +/+ and Popdc1 fs/fs rats. Prioritization of the top targets highlighted profound downregulation of BVES (POPDC1) and the membrane scaffold Ankyrin-G (Ank3) (n=5-7). (B) KEGG pathway enrichment of the proteome identified “Cardiac muscle contraction” and “Arrhythmogenic right ventricular cardiomyopathy” and various cardiomyopathies as the top disrupted pathways. (C) Immunofluorescence staining of the myocardium demonstrates the loss of both BVES and Ankyrin-G (both in red) in Popdc1 fs/fs rats. (D) RT-qPCR gene expression analyses in cardiac tissue demonstrated reduced Popdc1 but normalized AnkG mRNA (n=5-7). (E) Western blots confirm the reduction of cardiac POPDC1-Ankyrin-G-Nav1.5 axis and TREK-1 (KCNK2), concurrent with an upregulation of Kv4.3 and Cav1.2 in Popdc1 fs/fs rats (n=6). (F–G) Popdc1 fs/fs hearts exhibit markers of oxidative stress: elevated malondialdehyde (MDA; G) and reduced superoxide dismutase (SOD; H) activity (n=4). (F) Transmission electron microscopy (TEM) displays widened intercalated discs and myofibrillar disarray (red arrowhead) in Popdc1 fs/fs hearts (n=3). Data are mean ± SD. Statistical analyses conducted were unpaired Student’s t-test. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 respectively.

Article Snippet: Blots were probed with primary antibodies against Cav1.2 (1:1000, MA5-45389, Thermo Fisher Scientific), Kv4.3 (1:500, PA5-93292, Thermo Fisher Scientific), Nav1.5 (1:200, ASC-005, Alomone Labs), TREK-1/KCNK2 (1:1000, PA5-115452, Thermo Fisher Scientific), AnkG (1:1000, 27980-1-AP, Proteintech), BVES (1:1000, as above, Thermo Fisher Scientific), and GAPDH (1:10000, 60004-1-Ig, Proteintech).

Techniques: Membrane, Immunofluorescence, Staining, Quantitative RT-PCR, Gene Expression, Western Blot, Activity Assay, Transmission Assay, Electron Microscopy

Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A ) Schematic diagram showing the strategy for the drug screen. B ) THP-1 ASC-GFP monocytic cells without any treatment. C ) THP-1 ASC-GFP monocytic cells treated with LPS (1μg/ml) for 2 hours. D ) THP-1 ASC-GFP monocytic cells treated with LPS (1μg/ml) for 4 hours and Nigericin (10 μM) for 1 hour.

Article Snippet: The RAW264.7 cells (ATCC), RAW264.7-ASC (InvivoGen), RAW-DifluoTM mLC3 (InvivoGen), THP-1 (ATCC), THP-1-ASC-GFP (InvivoGen) were cultured in appropriate media containing required growth factors and antibiotics.

Techniques:

Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A ) Vehicle-treated or drug-treated, 20 μM, 16h ( B-P ) THP-1 ASC-GFP monocytic cells were treated with LPS (1μg/ml) for 4 hours. THP-1 ASC-GFP cells were pretreated with selected drugs for 16h, followed by washing with PBS and incubation with LPS (1μg/ml) for 2 hours.

Article Snippet: The RAW264.7 cells (ATCC), RAW264.7-ASC (InvivoGen), RAW-DifluoTM mLC3 (InvivoGen), THP-1 (ATCC), THP-1-ASC-GFP (InvivoGen) were cultured in appropriate media containing required growth factors and antibiotics.

Techniques: Incubation

Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A ) THP-1 ASC-GFP cells pretreated with vehicle or selected drugs for 16h, followed by washing with PBS and incubation with LPS (1μg/ml) for 2 hours, followed by incubation with Nigericin (10 μM) for 1 hour. ASC-puncta were visualized using fluorescent microscopy, with DAPI used to stain nuclei. B ) RAW-ASC cells pretreated with vehicle or selected drugs for 16h, followed by washing with PBS and incubation with LPS (1μg/ml) for 2 hours, followed by incubation with Nigericin (5 μM) for 1 hour. Cells were fixed and permeabilized and stained with anti-ASC antibody and Alexa-labeled secondary antibody. ASC-puncta were visualized using fluorescent microscopy, with DAPI used to stain nuclei.

Article Snippet: The RAW264.7 cells (ATCC), RAW264.7-ASC (InvivoGen), RAW-DifluoTM mLC3 (InvivoGen), THP-1 (ATCC), THP-1-ASC-GFP (InvivoGen) were cultured in appropriate media containing required growth factors and antibiotics.

Techniques: Incubation, Microscopy, Staining, Labeling

Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A, B ) Western blot analysis of basal or LPS-induced expression of various inflammasome components in THP-1 ASC-GFP cells pretreated with vehicle or selected drugs for 16h. C, D ) Western blot analysis of basal or LPS-induced expression of various inflammasome components in RAW- ASC cells pretreated with vehicle or selected drugs for 16h. E ) Age-matched (10-week-old) male WTC57BL6J mice fed with chow diet were i.p injected with saline or saquinavir. After 2 hours, mice were primed for inflammasome assembly by an I.P. injection of LPS (5μg/mouse). After 4h of LPS injection, the NLRP3 inflammasome assembly was induced by I.P. injection of ATP (0.5 mL of 30 mM, pH 7.0). The peritoneal cavity was lavaged with 5 mL sterile PBS, and IL-1β levels in peritoneal lavage were determined by ELISA (N = 6, mean ± SD for all groups, ∗∗p < 0.01 by two-tailed t-test). Mouse plasma was used in multiplex analysis to determine levels of F ) TNF-α, G ) IL-1β, H ) IL-17A, I ) IL-33, and J ) IL-15 (N=3, mean ± SD for all groups, ∗p < 0.05 by two-tailed t-test).

Article Snippet: The RAW264.7 cells (ATCC), RAW264.7-ASC (InvivoGen), RAW-DifluoTM mLC3 (InvivoGen), THP-1 (ATCC), THP-1-ASC-GFP (InvivoGen) were cultured in appropriate media containing required growth factors and antibiotics.

Techniques: Western Blot, Expressing, Injection, Saline, Sterility, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Clinical Proteomics, Multiplex Assay

Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A ) THP-1 ASC-GFP macrophages pretreated with vehicle or selected drugs for 16h, followed by washing with PBS and incubation with LPS (1μg/ml) for 2 hours, followed by incubation with Mitosox Red dye and fluorescent microscopy. DAPI staining is used to visualize nuclei.

Article Snippet: The RAW264.7 cells (ATCC), RAW264.7-ASC (InvivoGen), RAW-DifluoTM mLC3 (InvivoGen), THP-1 (ATCC), THP-1-ASC-GFP (InvivoGen) were cultured in appropriate media containing required growth factors and antibiotics.

Techniques: Incubation, Microscopy, Staining

Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A ) Vehicle-treated or selected drug-treated RAW-ASC cells were incubated with either saline or LPS (1μg/ml) for 2 hours, followed by indirect immunofluorescence using NF-kB antibody. B ) Quantification of NF-kB nuclear localization in RAW-ASC cells or C ) THP-1 macrophages, with bar graphs showing % of cells exhibiting nuclear localization of NF-kB (n > 50 for all, mean ± SD, * indicate p <0.05 **indicate p<0.01, ***indicate p<0.001 by t posttest). D, E ) Western blot analysis of either NLRP3, ASC, Caspase1, or IL-1β in RAW-ASC cells pretreated with either saline or selected drugs, followed by LPS treatment for 4 hours. Control cells were left untreated.

Article Snippet: RAW-ASC (Invivogen #raw-asc) were maintained in DMEM (Cleveland Clinic Media Core #11-500p) supplemented with 10% FBS, 1% Pen/Strep 5000u/mL, 100 μg/ml Blasticidin (Invivogen #ant-bl) and 100 μg/ml Normocin (Invivogen #ant-nr) RAW-DifluoTM mLC3 (Invivogen # awdf-mlc3) were maintained in DMEM supplemented with 10% FBS (Gibco), 1% pen-strep, 100 μg/ml Zeocin (Invivogen #ant-zn) and 100 μg/ml Normocin (Invivogen).

Techniques: Incubation, Saline, Immunofluorescence, Western Blot, Control

Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A ) RAW-ASC cells treated with control or drugs were incubated with Alexa-488-LPS for 2 hours, followed by washing and visualization under a fluorescent microscope.

Article Snippet: RAW-ASC (Invivogen #raw-asc) were maintained in DMEM (Cleveland Clinic Media Core #11-500p) supplemented with 10% FBS, 1% Pen/Strep 5000u/mL, 100 μg/ml Blasticidin (Invivogen #ant-bl) and 100 μg/ml Normocin (Invivogen #ant-nr) RAW-DifluoTM mLC3 (Invivogen # awdf-mlc3) were maintained in DMEM supplemented with 10% FBS (Gibco), 1% pen-strep, 100 μg/ml Zeocin (Invivogen #ant-zn) and 100 μg/ml Normocin (Invivogen).

Techniques: Control, Incubation, Microscopy

Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A ) THP-1 ASC-GFP cells pretreated with vehicle or selected drugs for 16h, followed by washing with PBS and incubation with LPS (1μg/ml) for 2 hours, followed by incubation with Nigericin (10 μM) for 1 hour. ASC-puncta were visualized using fluorescent microscopy, with DAPI used to stain nuclei. B ) RAW-ASC cells pretreated with vehicle or selected drugs for 16h, followed by washing with PBS and incubation with LPS (1μg/ml) for 2 hours, followed by incubation with Nigericin (5 μM) for 1 hour. Cells were fixed and permeabilized and stained with anti-ASC antibody and Alexa-labeled secondary antibody. ASC-puncta were visualized using fluorescent microscopy, with DAPI used to stain nuclei.

Article Snippet: RAW-ASC (Invivogen #raw-asc) were maintained in DMEM (Cleveland Clinic Media Core #11-500p) supplemented with 10% FBS, 1% Pen/Strep 5000u/mL, 100 μg/ml Blasticidin (Invivogen #ant-bl) and 100 μg/ml Normocin (Invivogen #ant-nr) RAW-DifluoTM mLC3 (Invivogen # awdf-mlc3) were maintained in DMEM supplemented with 10% FBS (Gibco), 1% pen-strep, 100 μg/ml Zeocin (Invivogen #ant-zn) and 100 μg/ml Normocin (Invivogen).

Techniques: Incubation, Microscopy, Staining, Labeling

Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A, B ) Western blot analysis of basal or LPS-induced expression of various inflammasome components in THP-1 ASC-GFP cells pretreated with vehicle or selected drugs for 16h. C, D ) Western blot analysis of basal or LPS-induced expression of various inflammasome components in RAW- ASC cells pretreated with vehicle or selected drugs for 16h. E ) Age-matched (10-week-old) male WTC57BL6J mice fed with chow diet were i.p injected with saline or saquinavir. After 2 hours, mice were primed for inflammasome assembly by an I.P. injection of LPS (5μg/mouse). After 4h of LPS injection, the NLRP3 inflammasome assembly was induced by I.P. injection of ATP (0.5 mL of 30 mM, pH 7.0). The peritoneal cavity was lavaged with 5 mL sterile PBS, and IL-1β levels in peritoneal lavage were determined by ELISA (N = 6, mean ± SD for all groups, ∗∗p < 0.01 by two-tailed t-test). Mouse plasma was used in multiplex analysis to determine levels of F ) TNF-α, G ) IL-1β, H ) IL-17A, I ) IL-33, and J ) IL-15 (N=3, mean ± SD for all groups, ∗p < 0.05 by two-tailed t-test).

Article Snippet: RAW-ASC (Invivogen #raw-asc) were maintained in DMEM (Cleveland Clinic Media Core #11-500p) supplemented with 10% FBS, 1% Pen/Strep 5000u/mL, 100 μg/ml Blasticidin (Invivogen #ant-bl) and 100 μg/ml Normocin (Invivogen #ant-nr) RAW-DifluoTM mLC3 (Invivogen # awdf-mlc3) were maintained in DMEM supplemented with 10% FBS (Gibco), 1% pen-strep, 100 μg/ml Zeocin (Invivogen #ant-zn) and 100 μg/ml Normocin (Invivogen).

Techniques: Western Blot, Expressing, Injection, Saline, Sterility, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Clinical Proteomics, Multiplex Assay