Journal: Arthritis Research & Therapy

Article Title: Upregulated endonuclease Regnase-1 suppresses osteoarthritis by forming a negative feedback loop of catabolic signaling in chondrocytes

doi: 10.1186/s13075-021-02485-z

Figure Lengend Snippet: SAA3 is a target of Regnase-1 in chondrocyte. a , b Number of genes modulated by Regnase-1 ( a ) and list of the top 13 genes that were most markedly regulated by Ad-shRegnase-1 ( b ). c Luciferase assay ( n = 4) in chondrocytes transfected with a reporter plasmid containing the SAA3 3′-UTR sequence. Chondrocytes were co-transfected with empty vector (EV), WT Regnase-1 (WT), or D141N Regnase-1 (D141N). d qRT-PCR analysis ( n = 6) of SAA family members in chondrocytes infected with Ad-Regnase-1 or Ad-shRegnase-1 for 36 h. e Quantitation of secreted SAAs by ELISA in chondrocytes treated with IL-1β (0.1 ng/ml, 36 h) or infected with the indicated MOI of Ad-C or Ad-SAA3 (36 h) ( n = 4). Means ± s.e.m. and one-way ANOVA with Bonferroni’s post hoc test. * p < 0.05, ** p < 0.005, *** p < 0.0005

Article Snippet: The SAA levels were analyzed using a mouse quantikine ELISA kit (R&D systems) according to the manufacturer’s protocol.

Techniques: Luciferase, Transfection, Plasmid Preparation, Sequencing, Quantitative RT-PCR, Infection, Quantitation Assay, Enzyme-linked Immunosorbent Assay

Journal: Critical Care Explorations

Article Title: Surfactant Protein D Influences Mortality During Abdominal Sepsis by Facilitating Escherichia coli Colonization in the Gut

doi: 10.1097/CCE.0000000000000699

Figure Lengend Snippet: Surfactant protein D (SPD) is synthesized by the gallbladder and promotes colonization of both the cecum and colon with Escherichia coli . A , SPD +/+ gut organs were harvested at baseline or post-cecal ligation and puncture (CLP) with Western blots performed for SPD or β -actin (loading control). Blots represent pooled samples, n = 2/group. Representative gel shown from three experiments. Lungs from SPD −/− and SPD +/+ mice were used as negative and positive controls, respectively. B , Gallbladder was isolated from SPD +/+ mice after CLP or sham surgery and from SPD −/− mice after CLP with Western blots performed for SPD or glyceraldehyde 3-phosphate dehydrogenase antibody (GAPDH; loading control). Blots represent pooled samples, n = 5–7/group. Lungs from SPD −/− mice and SPD +/+ were used as negative and positive controls, respectively. C , SPD −/− mice ( n = 9) were gavaged with recombinant surfactant protein D (rSPD), followed by gavage with green fluorescent protein (GFP)-labeled E. coli , and compared with SPD −/− mice gavaged only with GFP-labeled E. coli ( n = 9). After 24 hr, cecum and colon were harvested. GFP-labeled E. coli were then detected by culture (Mann-Whitney * p < 0.05, ** p < 0.01). CFU = colony forming units.

Article Snippet: SPD +/+ and SPD −/− mice were gavaged with 10 μL/g of ampicillin-resistant GFP-labeled E. coli (ATCC 25922GFP, 3.74 × 10 8 colony forming units/mL) with or without recombinant human surfactant protein D (rSPD) (Cat 1920-SP-050, R&D systems).

Techniques: Synthesized, Ligation, Western Blot, Control, Isolation, Recombinant, Labeling, MANN-WHITNEY