Journal: Respiratory physiology & neurobiology

Article Title: Paralemmin-3 augments lipopolysaccharide-induced acute lung injury with M1 macrophage polarization via the notch signaling pathway.

doi: 10.1016/j.resp.2023.104203

Figure Lengend Snippet: Fig. 5. Effect of PALM3 expression on Notch signaling pathway in AMs isolated from the BALF of each group. (A) Notch signaling associated molecules (NICD, RBP-J and IRAK2) mRNA and protein expression in in AMs isolated from the BALF of each group. (B) Co-immunoprecipitation assays of NICD and IRAK2 with RBP-J in AMs isolated from the BALF of each group. All data are expressed as means ± SD (n = 6). *P < 0.05 vs. normal group, and #P < 0.05 vs. empty vector group.

Article Snippet: Immunoprecipitation was performed with anti-rat-NICD antibody (1:50, #4147 monoclonal rabbit anti-rat, Cell Signaling Technology, Inc., Danvers, MA, USA), anti-ratRBP-J antibody (1:100, sc-271128, Santa Cruz) or normal rat IgG (1:100, sc-2026, Santa Cruz).

Techniques: Expressing, Isolation, Immunoprecipitation, Plasmid Preparation

Journal: Respiratory physiology & neurobiology

Article Title: Paralemmin-3 augments lipopolysaccharide-induced acute lung injury with M1 macrophage polarization via the notch signaling pathway.

doi: 10.1016/j.resp.2023.104203

Figure Lengend Snippet: Fig. 6. Involvement of Notch signaling pathway in PALM3-mediated M1 polarization. (A) The frequency of F4/80+CD16+ cells in AMs isolated from BALF were determined by flow cytometry with PE and FITC staining. All data are expressed as means ± SD (n = 6). *P < 0.05 vs. lenti.r-PALM3 + DMSO group, and #P < 0.05 vs. NS + DMSO group. (B) The mRNA levels of M1 specific genes (TNF-α, IL-1, CCr7 and iNOS) in AMs isolated from BALF were determined by qRT-PCR. All data are expressed as means ± SD (n = 6). *P < 0.05 vs. NS + DMSO group, and #P < 0.05 vs. lenti.r-PALM3 + DMSO group. (C) The mRNA levels of Notch signaling associated molecules (NICD, RBP-J and IRAK2) in AMs isolated from BALF were determined by qRT-PCR. All data are expressed as means ± SD (n = 6). *P < 0.05 vs. NS + DMSO group, and #P < 0.05 vs. lenti.r-PALM3 + DMSO group. (D) Co-immunoprecipitation assays of NICD and IRAK2 with RBP-J in AMs isolated from BALF. All data are expressed as means ± SD (n = 6). *P < 0.05 vs. NS + DMSO group, and #P < 0.05 vs. lenti.r-PALM3 + DMSO group.

Article Snippet: Immunoprecipitation was performed with anti-rat-NICD antibody (1:50, #4147 monoclonal rabbit anti-rat, Cell Signaling Technology, Inc., Danvers, MA, USA), anti-ratRBP-J antibody (1:100, sc-271128, Santa Cruz) or normal rat IgG (1:100, sc-2026, Santa Cruz).

Techniques: Isolation, Flow Cytometry, Staining, Quantitative RT-PCR, Immunoprecipitation

Journal: Respiratory physiology & neurobiology

Article Title: Paralemmin-3 augments lipopolysaccharide-induced acute lung injury with M1 macrophage polarization via the notch signaling pathway.

doi: 10.1016/j.resp.2023.104203

Figure Lengend Snippet: Fig. 7. Co-immunoprecipitation assays of NICD and RBP-J with PALM3 in AMs isolated from BALF. After rats were pretreated with lentiviral vectors, the AMs were collected from BALF on day 7 after successive LPS inhalations. The total cell lysates were immunoprecipitated with anti-NICD, anti-RBP-J or normal IgG antibodies. Then, the immunocomplexes were examined by western blot analysis using the indicated antibodies. All data are expressed as means ± SD (n = 6). *P < 0.05 vs. normal rat group.

Article Snippet: Immunoprecipitation was performed with anti-rat-NICD antibody (1:50, #4147 monoclonal rabbit anti-rat, Cell Signaling Technology, Inc., Danvers, MA, USA), anti-ratRBP-J antibody (1:100, sc-271128, Santa Cruz) or normal rat IgG (1:100, sc-2026, Santa Cruz).

Techniques: Immunoprecipitation, Isolation, Western Blot