Journal: European Journal of Medical Research

Article Title: Hydrogen gas inhalation ameliorates LPS-induced BPD by inhibiting inflammation via regulating the TLR4–NFκB–IL6/NLRP3 signaling pathway in the placenta

doi: 10.1186/s40001-024-01874-9

Figure Lengend Snippet: The sequence, product length and Ta opt of primers used for qRT-PCR analysis

Article Snippet: The PVDF membranes were incubated overnight with different antibodies at 4 °C: Rabbit monoclonal to NLRP3 (Abcam, ab263899, USA, 1:1000), Rabbit polyclonal to TLR4 (Abcam, ab217274, USA, 1:1000), Rabbit monoclonal to NF-kB p65 (phospho S536) (Abcam, ab76302, USA, 1:1000), Rabbit Anti-NFκB p65 antibody (Bioss, bs-0465R, CHINA, 1:800), Anti-CASP1 Rabbit polyclonal antibody (Merck, SAB5701133, CHINA, 1:1000), IκBα Rabbit pAb (ABclonal, A1187, CHINA, 1:1000), Phospho-IκBα-S32 Rabbit mAb (ABclonal, AP0707, CHINA, 1:1000) and β-actin Rabbit Monoclonal Antibody (Beyotime, AF5003, CHINA, 1:2000) and incubated with HRP-labeled Goat Anti-Rabbit IgG (H+L) (Beyotime, A0208, CHINA, 1:1000) for 2.5 h at least at room temperature. β-actin was selected as the internal reference protein, which is not only a protein encoded by the housekeeper gene, but also highly conserved and expressed relatively stable.

Techniques: Sequencing

Journal: European Journal of Medical Research

Article Title: Hydrogen gas inhalation ameliorates LPS-induced BPD by inhibiting inflammation via regulating the TLR4–NFκB–IL6/NLRP3 signaling pathway in the placenta

doi: 10.1186/s40001-024-01874-9

Figure Lengend Snippet: Hydrogen downregulated the placental TLR4–NFκB-IL6/NLRP3 inflammatory signaling pathway to rescue from LPS-induced BPD. A A representative image of Western blots for TLR4, IκBα, Phospho-IκBα (Ser32) ((p-IκBα(s32)), NF-κB p65, NF-kB p65 (phospho S536)(p-p65), NLRP3 and Casp1 in placenta from each group. β-actin was used to verify equivalent loading. Graphic representation of relative density of TLR4 ( B ), IκBα ( C ), p-IκBα(s32) ( D ), NF-κB p65 ( F ), p-p65(s536) ( G ), NLRP3 ( I ) and Casp1 ( J ) normalized to β-actin. Graphic representation of relative abundance of p-IκBα(s32)/IκBα ( E ) and p -p65/NF-κB p65 ( H ). Data were presented as mean ± SD of 8 samples from each group. * p < 0.05, ** p < 0.01 vs LPS group

Article Snippet: The PVDF membranes were incubated overnight with different antibodies at 4 °C: Rabbit monoclonal to NLRP3 (Abcam, ab263899, USA, 1:1000), Rabbit polyclonal to TLR4 (Abcam, ab217274, USA, 1:1000), Rabbit monoclonal to NF-kB p65 (phospho S536) (Abcam, ab76302, USA, 1:1000), Rabbit Anti-NFκB p65 antibody (Bioss, bs-0465R, CHINA, 1:800), Anti-CASP1 Rabbit polyclonal antibody (Merck, SAB5701133, CHINA, 1:1000), IκBα Rabbit pAb (ABclonal, A1187, CHINA, 1:1000), Phospho-IκBα-S32 Rabbit mAb (ABclonal, AP0707, CHINA, 1:1000) and β-actin Rabbit Monoclonal Antibody (Beyotime, AF5003, CHINA, 1:2000) and incubated with HRP-labeled Goat Anti-Rabbit IgG (H+L) (Beyotime, A0208, CHINA, 1:1000) for 2.5 h at least at room temperature. β-actin was selected as the internal reference protein, which is not only a protein encoded by the housekeeper gene, but also highly conserved and expressed relatively stable.

Techniques: Western Blot

Journal: European Journal of Medical Research

Article Title: Hydrogen gas inhalation ameliorates LPS-induced BPD by inhibiting inflammation via regulating the TLR4–NFκB–IL6/NLRP3 signaling pathway in the placenta

doi: 10.1186/s40001-024-01874-9

Figure Lengend Snippet: The relationships between TLR4–NFκB–IL6/NLRP3 and the protein levels of inflammatory cytokines/chemokines in placentas. A Relationship between TNF-α and placental p-IκBα(s32)/IκBα levels in all groups; B p-IκBα(s32)/IκBα vs p-p65/NF-κB p65; C p-p65/NF-κB p65 vs IL-6; D p-p65/NF-κB p65 vs CCL2; E p-p65/NF-κB p65 vs CXCL1; F p-p65/NF-κB p65 vs NLRP3; G NLRP3 vs Casp1; H – I Casp1 vs IL-18 and Casp1 vs IL-1β; n = 24, r = Pearson’s correlation coefficient

Article Snippet: The PVDF membranes were incubated overnight with different antibodies at 4 °C: Rabbit monoclonal to NLRP3 (Abcam, ab263899, USA, 1:1000), Rabbit polyclonal to TLR4 (Abcam, ab217274, USA, 1:1000), Rabbit monoclonal to NF-kB p65 (phospho S536) (Abcam, ab76302, USA, 1:1000), Rabbit Anti-NFκB p65 antibody (Bioss, bs-0465R, CHINA, 1:800), Anti-CASP1 Rabbit polyclonal antibody (Merck, SAB5701133, CHINA, 1:1000), IκBα Rabbit pAb (ABclonal, A1187, CHINA, 1:1000), Phospho-IκBα-S32 Rabbit mAb (ABclonal, AP0707, CHINA, 1:1000) and β-actin Rabbit Monoclonal Antibody (Beyotime, AF5003, CHINA, 1:2000) and incubated with HRP-labeled Goat Anti-Rabbit IgG (H+L) (Beyotime, A0208, CHINA, 1:1000) for 2.5 h at least at room temperature. β-actin was selected as the internal reference protein, which is not only a protein encoded by the housekeeper gene, but also highly conserved and expressed relatively stable.

Techniques: