Fig. 1
Spontaneously outgrown 32D CALR del52 cells are ruxolitinib (Ruxo) sensitive. a We performed growth assays with stably transduced 32D cells expressing empty vector (EV), WT CALR, del52 or the ins5 mutants (2 × 105 cells/ml). Outgrowth of a CALR del52 expressing clone has been observed after 6 to 10 days. b 32D EV, CALR WT, CALR del52, CALR ins5, and outgrown del52 cells were starved for 16 h; lysates were prepared and subjected to SDS-PAGE and Western blotting. Indicated antibodies have been used for immunostaining. c Outgrown 32D CALR del52 cells were treated overnight with 1 μM Ruxo, and reduction of STAT5 phosphorylation could be detected in Western blotting. d In a MTT assay, outgrown 32D CALR del52 cells were treated with 1 μM Ruxo for 48 h, and cell viability was evaluated. The experiment was performed in triplicates. SD is indicated. **P < 0.01 e Outgrown 32D del52 cells were grown in WEHI-free medium (2 × 105 cells/ml) supplemented with 1, 2, and 3 μM Ruxo or DMSO as control for up to 72 h. The cells were counted every 24 h. The experiment was performed in triplicates. SD is indicated