Fig. 1: EGF triggered ERK and RSK activation in the presence of ORF45. | Nature Communications

Fig. 1: EGF triggered ERK and RSK activation in the presence of ORF45.

From: A non-catalytic herpesviral protein reconfigures ERK-RSK signaling by targeting kinase docking systems in the host

Fig. 1

HEK293T (a) and HEK293T-ΔRSK1/2 (b) cells (‘HT-ORF45’) were treated with 100 ng/mL EGF and ERK (ppERK) and RSK (pS380) phosphorylation was monitored in Western-blots. Doxycycline treatment (DOX) was used to induce ORF45 expression (data for untreated and DOX treated cells are shown in black or red, respectively). Anti-panERK antibody was used as the load control for ppERK, and anti-tubulin antibody for pRSK Western-blots. Anti-phospho Western-blot signals were normalized to the Control signal (-: no DOX treatment) at 20 min for HEK293T cells and to the 7 min Control signal for HEK293T-ΔRSK1/2 cells. Error bars indicate SD of three independent experiments. Data points show the mean ± SD (N = 3). Western-blot panels show a representative set. c Mapping of RSK2 regions involved in the ORF45 mediated increase of ERK2-RSK2 binding. Different RSK2 constructs and ERK2 were tagged with the two different fragments of the split firefly luciferase enzyme and luminescence was normalized to the signal of the ERK2-RSK2 interaction (WT). WT: full-length RSK2, ΔD: RSK2 construct lacking the C-terminal tail containing the D-motif. Error bars show SD based on three independent experiments. (N = 3; Paired t-test, two-sided; NS not significant, *p < 0.05, **p < 0.01, ***p < 0.001). Source data are provided as Source data file.

Back to article page