radicicol  (Alomone Labs)

Journal: Epigenetics & Chromatin

Article Title: Pho dynamically interacts with Spt5 to facilitate transcriptional switches at the hsp70 locus

doi: 10.1186/s13072-017-0166-9

Figure Lengend Snippet: HSF recruitment to chromatin is necessary and sufficient for the dynamic protein–protein interaction between Pho and Spt5. a ChIP-qPCR measurements for the occupancy levels of HSF at the hsp70 locus in S2 DRSC cells upon HS (heat shock), radicicol, FP + HS (flavopiridol + heat shock) and Na Sal (Sodium Salicylate) treatment conditions, respectively. In case of radicicol treatment, the control condition was DMSO treatment. For FP + HS, the control condition was FP control and for Na Sal, the control condition was nuclease-free water. Distance of the location of the primers used from the hsp70 TSS is − 85 bp and has been depicted in the form of a cartoon below the data figure. b qRT-PCR measurements of hsp70 transcript levels in S2 DRSC cells under HS, radicicol, FP + HS and Na Sal treatment conditions, respectively. Distance of the location of the primers used from the hsp70 TSS is + 1492 bp and has been depicted in the form of a cartoon below the data figure. c co-IP assays of S2 DRSC cells transiently transfected with plasmids expressing FLAG-tagged Spt5 (FLAG-Spt5) and HA-tagged Pho (HA-Pho). The S2 DRSC cells were subjected to HS, radicicol, FP + HS and Na Sal, respectively; cell lysates were used for pull-downs with an anti-FLAG antibody and later probed by Western blot with an anti-HA antibody. d Quantification of the co-IP assays in c . Details of the procedure used for quantification can be found in Materials and Methods. e co-IP assays of S2 DRSC cells transiently transfected with plasmids expressing FLAG-tagged Spt5 (FLAG-Spt5) and HA-tagged Pho (HA-Pho). Cells were either treated with HSF or LacZ RNAi for 4 days, then either maintained at control conditions or heat shocked, cell lysates were prepared and used for pull-downs using an anti-FLAG antibody and later probed by Western blot using an anti-HA antibody. f qRT-PCR measurements of hsp70 transcript levels in S2 DRSC cells under HSF and LacZ RNAi conditions. Distance of the location of the primers used from the hsp70 TSS is + 1492 bp and has been depicted in the form of a cartoon below the data figure. Data information: In a and b , data are presented as mean ± SEM for n = 2, while in e , data are presented as mean ± SEM for n = 3

Article Snippet: Chemicals and antibodiesThe following chemicals were used in this study: flavopiridol (F3055, Sigma-Aldrich), radicicol (R-370, Alomone), sodium salicylate (S3007, Sigma-Aldrich), copper sulphate (102790, Merck).

Techniques: Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Co-Immunoprecipitation Assay, Transfection, Expressing, Western Blot

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Trithorax requires Hsp90 for maintenance of active chromatin at sites of gene expression

doi: 10.1073/pnas.0809669106

Figure Lengend Snippet: Hsp90 is required for the stability of the Trx protein. ( A ) In Drosophila Kc cells, inhibition of Hsp90 with radicicol (Radic) resulted in depletion of Trx protein. After 4 h of Radic treatment with 20 μM (lane 2) and 40 μM (lane 4), Trx

Article Snippet: A total of 3 × 106 /ml cells per well in a 6-well plate were seeded 1 day before drug treatment and treated with different concentrations (3, 5, 10, 20, 40, and 80 μM) of radicicol dissolved in DMSO (Alomone labs) ( A and data not shown) for 4 h. The Kc cells treated with equivalent amounts of DMSO vehicle were used as mock-treated cells.

Techniques: Inhibition