Journal: Science advances

Article Title: Targeting pyroptosis with nanoparticles to alleviate neuroinflammatory for preventing secondary damage following traumatic brain injury.

doi: 10.1126/sciadv.adj4260

Figure Lengend Snippet: Fig. 4. Inhibitory effect of C-β-LG/DSF on pyroptosis after TBI. (A) The schematic diagram illustrates the inhibition of pyroptosis by DSF. After the uptake of C-β-LG/DSF by cells, DSF inhibits the aggregation of GSDMD N-terminal pores on the cell membrane surface. Furthermore, experimental results demonstrated that the expression of GSDMD, GSDMD-N, caspase-1, and other related proteins was reduced after treatment with C-β-LG/DSF. (B) Immunoblotting of GSDMD, GSDMD-N, and caspase-1, caspase-1 p10, caspase-1 p20 in neuron-ICR cells with sham, TBI, DSF, and C-β-LG/DSF for 24 hours. (C to E) Levels of (C) IL-1β, (D) IL-18, and (E) LDH in injured neuron-ICR as detected by ELISA 12 hours after TBI. (F) Immunohistochemical staining was used to observe the expression of GSDMD in injured tissues of TBI model mice 3 days after different drugs treatment. Scale bars, 50 μm. (G) Quantitative analysis of GSDMD-positive cells. (H to J) Levels of (H) IL-1β, (I) IL-18, and (J) LDH in injured tissues as detected by ELISA 3 days after TBI. (K) Immunoblotting of GSDMD, GSDMD-N, caspase-1, caspase-1 p20, and caspase-1 p10 in injured tissue with sham, TBI, DSF, β-LG/DSF, and C-β-LG/ DSF for 24 hours. Data were expressed as means ± SD (n = 5). For (C) to (E) and (G) to (J), statistical analysis was calculated via one-way ANOVA test. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

Article Snippet: Primary antibodies used for immunoblot were listed as follows: anti– caspase- 1 p20 (Santa Cruz Biotechnology, sc- 398715; 1:1000), anti– cleaved GSDMD (Cell Signaling Technology, #10137S; 1:1000), anti–caspase- 1 p10 (GeneTex, GTX134551; 1:1000), and anti- GAPDH (Proteintech, 60004- 1- Ig; 1:1000).

Techniques: Inhibition, Membrane, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining

Journal:

Article Title: Minocycline inhibits caspase-independent and -dependent mitochondrial cell death pathways in models of Huntington's disease

doi: 10.1073/pnas.1832501100

Figure Lengend Snippet: Minocycline inhibits AIF, Smac, and cytochrome c release, caspase activation, and Bid cleavage in R6/2 mice. Cytosolic fractions from 10.5-week-old R6/2 mice or wild-type mice treated with i.p. injections of saline or minocycline were analyzed by Western blot with AIF, Smac, and cytochrome c antibodies. Brain lysates from above mice were separated by SDS/PAGE and probed with caspase-1, -9, -3, and -8 and Bid antibodies. The same blot was reprobed with a β-actin antibody. Densitometry was performed to quantify each lane (n = 3 mice per condition, *, P < 0.05).

Article Snippet: Caspase-8 and -3 antibodies were purchased from PharMingen, caspase-9 antibody from Cell Signaling Technology (Beverly, MA), caspase-1 antibody from Santa Cruz Biotechnology, BID antibody from R & D Systems, β-actin antibody from Sigma, and histone H2A antibody from MBL (Watertown, MA).

Techniques: Activation Assay, Western Blot, SDS Page

Journal: Journal of Nanobiotechnology

Article Title: Manganese-enriched nanoboron agent amplifies BNCT efficacy via pyroptosis-mediated immune activation and STING pathway synergy​

doi: 10.1186/s12951-025-03916-8

Figure Lengend Snippet: Pyroptosis Induced by BSA- 10 BPA-MnO 2 after BNCT. ( a ) Fluorescent images of ROS in B16F10 cells after different treatments, detected by DCFH-DA staining. Scale bar: 150 μm. ( b ) Cell morphology images after different treatments. In the BSA- 10 BPA-MnO 2 + N group, cell swelling and the appearance of large bubbles (blue arrows, one of the most prominent features of pyroptosis) are observed. Scale bar: 50 μm. ( c ) Western blot analysis of caspase-1 (CAS-1) and cleaved caspase-1 (cleaved-CAS-1). ( d ) Immunofluorescent staining of the GSDMD protein N-terminal fragment (GSDMD-N). Scale bar: 100 μm. ( e ) Relative LDH release, n = 4. Data are presented as mean ± SD. * p < 0.5, **** p < 0.0001

Article Snippet: After blocking, membranes were probed with primary antibodies against caspase-1 and cleaved caspase-1 (MedChemExpress, China), followed by HRP-conjugated secondary antibodies.

Techniques: Staining, Western Blot