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Mol Cell. Author manuscript; available in PMC 2021 Mar 19.
Published in final edited form as:
Mol Cell. 2020 Mar 19; 77(6): 1294–1306.e5.
Published online 2020 Feb 4. doi: 10.1016/j.molcel.2020.01.009

Figure 6.

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SPHK1 is an SFMBT1 direct target gene in ccRCC.

(A) ChIP-seq binding peaks on SPHK1 for SFMBT1, HIF2α, HIF1β, H3K4me1, H3K4me3, H3K27ac and H3K27me3.

(B) SPHK1 protein level in cells transfected with control siRNA (Ctrl) or SFMBT1 si-1 and si-3.

(C-E) Immunoblots for lysates (C), cell proliferation assays (D) and representative anchorage-independent growth assays (E) in cells infected with lentivirus encoding either empty vector (EV), HA-SFMBT1 or P651A. * EV/SFMBT1 vs P651A; **, P<0.01. The values listed below the blots (C) indicate the relative SPHK1 protein levels with Tubulin normalization.

(F-H) Immunoblots for lysates (F), cell proliferation assays (G) and representative anchorage-independent growth assays (H) in cells infected with lentivirus encoding either empty vector (EV), P651A with or without PF543 treatment. * P651A vs Ctrl; # P651A+PF543 vs Ctrl. */#, P<0.05; **/# #, P<0.01.

(I-K) Immunoblots for lysates (I), cell proliferation assays (J) and representative anchorage-independent growth assays (K) in cells transduced with lentivirus expressing either SFMBT1 shRNA control (Ctrl) or SFMBT1 sh1, followed by infection with lentivirus encoding either empty vector (EV) or v5-SPHK1. * EV+shSFMBT1 sh1 vs EV; # v5-SPHK1+SFMBT1 sh1 vs EV. */#, P<0.05; **/# #, P<0.01.

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