(A) The histograms show the MFI for PD-L1 on K562 cells (transfected with
empty vector or JAK2V617F vector). One representative experiment of three
experiments with a comparable pattern is shown. The analysis was done on
GFP+ sorted cells within 3 days after transfection.
(B) The bar diagram displays the fold change of PD-L1 expression (flow
cytometry) on K562 cells transfected with empty vector or with JAK2V617F. The
data are pooled from 4 independent experiments (n=12 per group).
(C) The bar diagram displays the fold change of PD-L1 expression (flow
cytometry) on K562 JAK2V617F cells that were exposed to different
concentrations of the JAK2 inhibitor SD-1029. Pooled data from two independent experiments
(n=6 at each concentration).
(D) The bar diagram displays the fold change of PD-L1 expression (flow
cytometry) for the JAK2V617F-positive cell line UKE-1 treated with the JAK2
inhibitor SD-1029 (n=7 at each concentration).
(E) The Western blots display STAT3 total protein, β-actin and
phospho-STAT3 in UKE-1 cells being treated with the JAK2 inhibitor SD-1029. The blots are
representative of three independent experiments.
(F) The bar diagram indicates the ratio of pSTAT3/STAT3/β-actin
(normalized to 1 in the condition without JAK2 inhibitor) for the cells described in (E).
Pooled data from three replicates (n=3 for each concentration).
(G) The bar diagram displays the fold change of PD-L1 expression (flow
cytometry) for JAK2V617F positive cell line SET-2 treated with ruxolitinib.
Pooled data from three independent experiments (concentration 0 – 0.5 μM:
n=12, concentration 1 and 2 μM: n=6).
(H) The Western blots display STAT3, β-actin, and phospho-STAT3 in
SET-2 cells being treated with ruxolitinib. The blots are representative of three
independent experiments.
(I) The bar diagram indicates the pSTAT3/STAT3/β-actin ratio for
cells described in (H). Pooled data from three independent experiments (n=3 for
each concentration).
(J) The bar diagram displays the fold change of PD-L1 expression (flow
cytometry) for JAK2V617F positive cell line MUTZ-8 that was treated with the
JAK2 inhibitor SD-1029 (n=3 for each concentration).
(K) The Western blots display STAT3 total protein, β-actin and
phospho-STAT3 in MUTZ-8 cells being treated with the JAK2 inhibitor SD-1029. The blots are
representative of three independent experiments.
(L) The bar diagram indicates the ratio of pSTAT3/STAT3/β-actin for
cells described in (K). Pooled data from three independent experiments (n=3 for
each concentration).
(M) The bar diagram indicates the relative luminescence activity of K562
cells (containing empty vector or JAK2V617F) which were left untransfected or
transfected with either a promoterless luciferase reporter-vector pgl4.13 or a reporter
vector containing the PD-L1 promoter. Pooled data from three technical replicates
(n=6 for each condition).
(N) The bar diagram indicates the relative luciferase activity of K562
JAK2V617F cells transfected with either the promoterless luciferase reporter
vector pgl4.13 or the reporter vector containing the PD-L1 promoter and treated with the
JAK2-inhibitor SD-1029. Pooled data from three independent experiments (n=6 for
each condition).
(O) The bar diagram indicates the relative luminescence activity of K562
JAK2V617F cells transfected with either the promoterless luciferase reporter
vector pgl4.13 or the reporter vector containing the PD-L1 promoter and treated with the
STAT3 inhibitor S3I-201. Pooled data from three independent experiments (n=6 for
each condition).