mouse il 18 duoset elisa kit  (R&D Systems)

Journal: Neural Regeneration Research

Article Title: Phosphodiesterase 4 regulates pyroptosis in subarachnoid hemorrhage

doi: 10.4103/NRR.NRR-D-24-01381

Figure Lengend Snippet: Activation of PDE4 induces neuronal pyroptosis 24 hours after SAH. (A) Representative TEM images of neurons. Red arrows: membrane pores. (B) Confocal IF staining of PDE4 (red) with neurons (NeuN, green) and proportion of PDE4-positive neurons. (C) Confocal IF staining of NLRP3 (red) with neurons (NeuN, green) and proportion of NLRP3-positive neurons. (D) Confocal IF staining of caspase-1 (red) with neurons (NeuN, green) and proportion of caspase-1-positive neurons. (E) Confocal IF staining of GSDMD (red) with neurons (NeuN, green) and proportion of GSDMD-positive neurons. Scale bars: 2 μm in A and 20 μm in B–E. (F) Representative western blot bands and relative protein expression of NLRP3, ASC, cleaved caspase-1, and GSDMD-N. (G) The levels of IL-1β and IL-18 in the cerebral cortex of mice detected by enzyme-linked immunosorbent assay. Data are presented as mean ± SD; n = 6 per group. ** P < 0.01, *** P < 0.001, **** P < 0.0001 (one-way analysis of variance with Tukey’s post hoc test). ASC: Apoptosis-associated speck-like protein containing a caspase recruitment domain; Caspase-1: cysteinyl aspartate specific proteinase-1; ETZ: etazolate; GSDMD: gasderminD; IF: immunofluorescence; IL: interleukin; NLRP3: nucleotide-binding oligomerization domain-like receptor pyrin domain containing 3; PDE4: phosphodiesterase 4; SAH: subarachnoid hemorrhage; Sham: sham surgery; TEM: transmission electron microscopy.

Article Snippet: The inflammatory factors of the brain cortex and primary neurons were measured using a mouse interleukin (IL)-1β ELISA kit (RK04878, Abclonal, Wuhan, China) and a mouse IL-18 ELISA kit (RK00104, Abclonal) following the manufacturer’s instructions.

Techniques: Activation Assay, Membrane, Staining, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Binding Assay, Transmission Assay, Electron Microscopy

Journal: Neural Regeneration Research

Article Title: Phosphodiesterase 4 regulates pyroptosis in subarachnoid hemorrhage

doi: 10.4103/NRR.NRR-D-24-01381

Figure Lengend Snippet: Activation of PDE4 induces pyroptosis of primary neurons 24 hours after OxyHb induction. (A) Confocal IF staining of PDE4 (red) with primary neurons (MAP2, green) and PDE4 relative fluorescence intensity. (B) Confocal IF staining of NLRP3 (red) with primary neurons (MAP2, green) and NLRP3 relative fluorescence intensity. (C) Confocal IF staining of caspase-1 (red) with primary neurons (MAP2, green) and caspase-1 relative fluorescence intensity. (D) Confocal IF staining of GSDMD (red) with primary neurons (MAP2, green) and GSDMD relative fluorescence intensity. Scale bars: 20 μm in A–D. (E) Representative western blot bands and relative protein expression of PDE4, NLRP3, ASC, cleaved caspase-1, and GSDMD-N. (F) The levels of IL-1β and IL-18 in primary neurons detected by enzyme-linked immunosorbent assay. Data are presented as mean ± SD; n = 6 per group. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 (one-way analysis of variance with Tukey’s post hoc test or the Brown–Forsythe test). ASC: Apoptosis-associated speck-like protein containing a caspase recruitment domain; Caspase-1: cysteinyl aspartate specific proteinase-1; cells: primary neurons; ETZ: etazolate; GSDMD: gasderminD; IF: immunofluorescence; IL: interleukin; NLRP3: nucleotide-binding oligomerization domain-like receptor pyrin domain containing 3; OxyHb: oxyhemoglobin; PDE4: phosphodiesterase 4.

Article Snippet: The inflammatory factors of the brain cortex and primary neurons were measured using a mouse interleukin (IL)-1β ELISA kit (RK04878, Abclonal, Wuhan, China) and a mouse IL-18 ELISA kit (RK00104, Abclonal) following the manufacturer’s instructions.

Techniques: Activation Assay, Staining, Fluorescence, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Binding Assay

Journal: Neural Regeneration Research

Article Title: Phosphodiesterase 4 regulates pyroptosis in subarachnoid hemorrhage

doi: 10.4103/NRR.NRR-D-24-01381

Figure Lengend Snippet: Knockdown of endogenous PDE4 improved EBI and neuronal pyroptosis 24 hours after SAH. (A) Confocal IF staining of Cy3-conjugated PDE4 siRNA (red) with neurons (NeuN, green) and proportion of PDE4 siRNA-positive neurons. Scale bars: 50 μm. (B) The modified Garcia score, balance beam test, Rotarod test, and brain water content of all groups. (C) Representative western blot bands and relative protein expression of PDE4, NLRP3, ASC, cleaved caspase-1, and GSDMD-N. (D) The levels of IL-1β and IL-18 in the cerebral cortex of mice detected by enzyme-linked immunosorbent assay. Data are presented as mean ± SD; n = 6 per group; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 (one-way analysis of variance with Tukey’s post hoc test or the Brown–Forsythe test). ASC: Apoptosis-associated speck-like protein containing a caspase recruitment domain; Caspase-1: cysteinyl aspartate specific proteinase-1; EBI: early brain injury; GSDMD: gasderminD; IF: immunofluorescence; IL: interleukin; NLRP3: nucleotide-binding oligomerization domain-like receptor pyrin domain containing 3; PDE4: phosphodiesterase 4; SAH: subarachnoid hemorrhage; Scr: scrambled; Sham: sham surgery; siRNA: small interfering RNA.

Article Snippet: The inflammatory factors of the brain cortex and primary neurons were measured using a mouse interleukin (IL)-1β ELISA kit (RK04878, Abclonal, Wuhan, China) and a mouse IL-18 ELISA kit (RK00104, Abclonal) following the manufacturer’s instructions.

Techniques: Knockdown, Staining, Modification, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Binding Assay, Small Interfering RNA

Journal: Neural Regeneration Research

Article Title: Phosphodiesterase 4 regulates pyroptosis in subarachnoid hemorrhage

doi: 10.4103/NRR.NRR-D-24-01381

Figure Lengend Snippet: Specific activation of NLRP3 reversed the neuroprotective effect of PDE4 inhibition 24 hours after SAH. (A) The modified Garcia score, balance beam test, Rotarod test, and brain water content of all groups. (B) Representative western blot bands and relative protein expression of PDE4, NLRP3, ASC, cleaved caspase-1, and GSDMD-N in the cerebral cortex of mice. (C) The levels of IL-1β and IL-18 in the cerebral cortex of mice detected by enzyme-linked immunosorbent assay. Data are presented as mean ± SD; n = 6 per group; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 (one-way analysis of variance with Tukey’s post hoc test or the Brown–Forsythe test). ASC: Apoptosis-associated speck-like protein containing a caspase recruitment domain; Caspase-1: cysteinyl aspartate specific proteinase-1; ETZ: etazolate; GSDMD: gasderminD; IL: interleukin; NIG: nigericin; NLRP3: nucleotide-binding oligomerization domain-like receptor pyrin domain containing 3; ns: not significant; PDE4: phosphodiesterase 4; SAH: subarachnoid hemorrhage; Sham: sham surgery.

Article Snippet: The inflammatory factors of the brain cortex and primary neurons were measured using a mouse interleukin (IL)-1β ELISA kit (RK04878, Abclonal, Wuhan, China) and a mouse IL-18 ELISA kit (RK00104, Abclonal) following the manufacturer’s instructions.

Techniques: Activation Assay, Inhibition, Modification, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Binding Assay