Fig. 4

Pho preferentially interacts with Spt5 and dSfmbt during the active and silencing phase, respectively, of the heat shock response. a, c co-IP assays of S2 DRSC cells transiently transfected with plasmids expressing FLAG-tagged dSfmbt or Spt5 (FLAG-dSfmbt, FLAG-Spt5) and HA-tagged Pho (HA-Pho). The S2 DRSC cells were subjected to a heat shock and then allowed to recover from the heat shock according to the experimental scheme depicted in Fig. 1a. At the indicated time points in the experimental scheme, cell lysates were prepared and were used for pull-downs using an anti-FLAG antibody. They were later probed by Western blot using an anti-HA antibody. b, d Quantification of the co-IP assays in A and C. The control line represents cells that were maintained at 25 °C for the entire duration of the time course detailed in Fig. 1a. Details of the procedure used for quantification can be found in Materials and Methods. Data information: In b and d, data are presented as mean ± SEM for n = 2