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Image Search Results
Journal: PLoS Biology
Article Title: Hypoxia-Inducible Factor-2α Is an Essential Catabolic Regulator of Inflammatory Rheumatoid Arthritis
doi: 10.1371/journal.pbio.1001881
Figure Lengend Snippet: (A and B) Typical immunofluorescence microscopy images of DAPI, HIF-2α, and FLS markers (vimentin and CD55) or the macrophage marker CD68 in human RA synovium (A) and mouse CIA synovium (B). (C, Left) Primary culture of total synovial cells isolated from DBA/1J mice. DC, dendritic cells; MΦ, macrophages. (Center) Typical immunofluorescence microscopy images of HIF-2α, DAPI, and vimentin or CD68. (Right) The percentage of FLS and macrophages positive for HIF-2α staining was determined from six microscopic fields ( n = 4). (D) Raw264.7 cells were treated with LPS (50 ng/ml) or TNFα (50 ng/ml) for 24 h. mRNA levels were detected by RT-PCR analysis ( n = 6). (E) Primary cultured FLS were treated with IL1β (1 ng/ml), IL6 (100 ng/ml), IL17 (10 ng/ml), or TNFα (100 ng/ml) for 24 h. mRNA levels of HIF-1α and HIF-2α were quantified by qRT-PCR ( n = 6). (F) FLS were treated with PD98059 (PD; 20 µM) to inhibit ERK, SB203580 (SB; 20 µM) to inhibit p38 MAP kinase, SP600125 (SP; 20 µM) to inhibit JNK, or the indicated concentration (µM) of BAY 11-7085 (BAY) to inhibit NF-κB. The cells were exposed to IL1β or TNFα for 24 h, and HIF-2α mRNA levels were quantified ( n = 6). (G) Mouse CIA synovium was stained for the hypoxia marker pimonidazole (upper). Primary cultured FLS were maintained under hypoxic conditions or were infected with Ad- Epas1 at an MOI of 800 for 24 h. HIF-1α and HIF-2α proteins were detected by Western blotting (lower). Values are presented as means ± SEM (* p <0.01, ** p <0.005, *** p <0.001). Scale bar, 50 µm.
Article Snippet: For double-immunofluorescence labeling of human and mouse joint tissues, the following primary antibodies were used: rabbit anti–HIF-2α (Novus Biologicals for human tissues and Santa Cruz for mouse tissues), rabbit anti-MMP3 (Abcam), mouse anti-MMP13 (Calbiochem, for human synovia), rabbit anti-MMP13 (Abcam, for mouse synovia), goat anti-IL6 (R&D Systems),
Techniques: Immunofluorescence, Microscopy, Marker, Isolation, Staining, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Quantitative RT-PCR, Concentration Assay, Infection, Western Blot
Journal: International Journal of Molecular Sciences
Article Title: Urinary Tract Tumor Organoids Reveal Eminent Differences in Drug Sensitivities When Compared to 2-Dimensional Culture Systems
doi: 10.3390/ijms23116305
Figure Lengend Snippet: Reagents for immunofluorescence staining.
Article Snippet:
Techniques: Immunofluorescence, Staining