Journal: Cell reports

Article Title: High-dimensional profiling clusters asthma severity by lymphoid and non-lymphoid status

doi: 10.1016/j.celrep.2021.108974

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-Human IFN-g (B27)-158Gd , Fluidigm , Cat# 3158017B.

Techniques: Purification, Antibody Labeling, Flow Cytometry, Software, Staining, Blocking Assay

Journal: Molecules

Article Title: Stemodane Diterpenes and Diterpenoids: Isolation, Structure Elucidation, Biogenesis, Biosynthesis, Biological Activity, Biotransformations, Metabolites and Derivatives Biological Activity, Rearrangements

doi: 10.3390/molecules26092761

Figure Lengend Snippet: Values of the minimal inhibitory concentration (MIC) (μg/mL) of (+)-stemodin ( 1 ) and (+)-stemodinoside B ( 15 ) [ 6 ]. ND = not determined.

Article Snippet: Escherichia coli (ATCC 27) , 512 , ≥1024.

Techniques: Concentration Assay

Journal: The Journal of Clinical Investigation

Article Title: HLA-B27–mediated activation of TNAP phosphatase promotes pathogenic syndesmophyte formation in ankylosing spondylitis

doi: 10.1172/JCI125212

Figure Lengend Snippet: (A) ARS staining of mineralization in AS MSCs transduced with shHLA-B or shCtrl under osteogenic induction with quantification (B). (C) Immunoblot analyses showing the expression of RARB and TNAP in AS MSCs expressing shHLA-B or shCtrl at day 7 under osteogenic induction. (D) Immunoblot showing HLA-B27 expressions of control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. (E) ARS staining of control MSCs transduced with control lentiviral vector (pLAS2w) or a vector expressing HLA-B27 (pLAS2w-HLA-B27) with quantification (F). (G) Immunoblot analyses showing RARB and TNAP expressions in control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. All experiments done in the AS patient group and the control group are from AS MSCs (A1, A2, and A3) and control MSCs (C1, C2, and C3), respectively, with at least 2–3 experimental repeats. Data are the mean ± SEM. ***P < 0.001; ****P < 0.0001 by 2-tailed Student’s t test (2 groups) or 1-way ANOVA, followed by Tukey’s HSD test. Representative images from AS (A1) MSCs are shown in C. Scale bars: 200 μm (A and E).

Article Snippet: The primary antibodies were mouse anti–active dephosphorylated β-catenin (clone 8E7; 1:500; Merck Millipore), rabbit anti–phosphorylated (serine 463/465) Smad 1/5/8 (clone 41D10; 1:250; Cell Signaling Technology), rabbit anti-TNAP (clone EPR4477; 1:500; Abcam), rabbit anti-RARB (catalog ab53161; 1:500; Abcam), goat anti-LEF1 (clone N-17; 1:500; Santa Cruz Biotechnology), rabbit anti-IRE1 (catalog ab37073; 1:1000; Abcam), rabbit anti–phosphorylated (serine 724) IRE1 (catalog ab4818; 1:500; Abcam), rabbit anti-XBP1 (catalog ab37152; 1:2000; Abcam), anti–HLA-B27 (clone HLA-ABC-m3; 1:300; Origene), anti–HLA-B7 [clone EPR2623( 2 ); 1:500; Abcam], rabbit anti-GAPDH (catalog GTX100118; 1:10,000; GeneTex), and mouse anti–β-actin (catalog MA5-15739; 1:10,000; Thermo Fisher Scientific).

Techniques: Staining, Transduction, Western Blot, Expressing, Plasmid Preparation

Journal: The Journal of Clinical Investigation

Article Title: HLA-B27–mediated activation of TNAP phosphatase promotes pathogenic syndesmophyte formation in ankylosing spondylitis

doi: 10.1172/JCI125212

Figure Lengend Snippet: (A) Unfolded or misfolded HCs were immunoprecipitated (IP) from lysates of AS MSCs and control MSCs with HC10 antibody, followed by immunoblotting with HLA-B27 and anti-BiP antibodies. In AS MSCs, monomers of HLA-B27 HCs and dimers of disulfide-linked HLA-B27 HCs are indicated by an arrow and an arrowhead, respectively. (B and C) Immunoblot showing the expressions of p-IRE1 (B) and sXBP1 (C) protein in AS and control MSCs at day 7 under osteogenic induction. (D) Immunoblot showing the expressions of p-IRE1 and sXBP1 in AS MSCs transduced with shHLA-B or shCtrl. (E) Immunoblot showing the expressions of p-IRE1 and sXBP1 protein in control MSCs transduced with pLAS2w-HLA-B27 or pLAS2w. (F) ChIP assay showing sXBP1 binding at fragments (Frags) 1, 2, 3, 6, and 7 within 1400 bp upstream of the RARB transcriptional start site in AS MSCs. RARB 3′-untranslated region (3′UTR) was used as the negative control locus. (G) Immunoblot showing the expressions of RARB and TNAP in AS MSCs expressing shXBP1 or shCtrl. (H) ARS staining of mineralization in AS MSCs transduced with shXBP1 or shCtrl under osteogenic induction with quantification (I). All experiments done in the AS patient group and control group are from AS MSCs (A1, A2, and A3) and control MSCs (C1, C2, and C3), respectively, with at least 2–3 experimental repeats. Data are the mean ± SEM. **P < 0.05; ***P < 0.001; ****P < 0.0001 by 2-tailed Student’s t test (2 groups) or ****P < 0.0001 by 1-way ANOVA, followed by Tukey’s HSD test. Representative immunoblots from AS (A1) MSCs are shown in G.

Article Snippet: The primary antibodies were mouse anti–active dephosphorylated β-catenin (clone 8E7; 1:500; Merck Millipore), rabbit anti–phosphorylated (serine 463/465) Smad 1/5/8 (clone 41D10; 1:250; Cell Signaling Technology), rabbit anti-TNAP (clone EPR4477; 1:500; Abcam), rabbit anti-RARB (catalog ab53161; 1:500; Abcam), goat anti-LEF1 (clone N-17; 1:500; Santa Cruz Biotechnology), rabbit anti-IRE1 (catalog ab37073; 1:1000; Abcam), rabbit anti–phosphorylated (serine 724) IRE1 (catalog ab4818; 1:500; Abcam), rabbit anti-XBP1 (catalog ab37152; 1:2000; Abcam), anti–HLA-B27 (clone HLA-ABC-m3; 1:300; Origene), anti–HLA-B7 [clone EPR2623( 2 ); 1:500; Abcam], rabbit anti-GAPDH (catalog GTX100118; 1:10,000; GeneTex), and mouse anti–β-actin (catalog MA5-15739; 1:10,000; Thermo Fisher Scientific).

Techniques: Immunoprecipitation, Western Blot, Transduction, Binding Assay, Negative Control, Expressing, Staining