raw asc  (InvivoGen)

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