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. 2024 Feb 16;15:1442. doi: 10.1038/s41467-024-45852-5

Fig. 7. CDK6 and CAD inhibitors block KSHV pathogenesis.

Fig. 7

A HOKs infected with KSHV (MOI = 30) for 48 h were treated with vehicle or Palbociclib for 4 h. WCLs were precipitated with anti-CAD. Precipitated proteins were analyzed by immunoblotting with the indicated antibodies. B BCBL-1 cells were treated with Palbociclib (50 nM). WCLs were precipitated with anti-CAD. Precipitated proteins were analyzed by immunoblotting with the indicated antibodies. C HOKs infected with KSHV (MOI = 30) for 48 h were treated with vehicle or Palbociclib for 4 h. WCLs were analyzed by 2DGE. D LECs infected with KSHV (MOI = 10) for 24 h were treated with DON (30 µM) or Palbociclib (2 µM) for 48 h. 72 h post infection, RT-qPCR analysis of the indicated mRNAs was performed. E LECs infected with KSHV (MOI = 10) for 24 h were treated with DON (30 µM) or Palbociclib (2 µM). Lactate in the media was determined at the indicated hours. F RT-qPCR analysis of viral mRNAs in LECs infected with KSHV (MOI = 10) for 24 h and then treated with Palbociclib (L: 0.4 µM, H: 2 µM) for 24 h. ND: undetected. G Viral titers in the media of the infected LECs in (E). H Viability of BC-3, BCBL-1, BJAB, and JSC-1 cells in the presence of DON. I Viability of BC-3, BCBL-1, BJAB, and JSC-1 cells in the presence of Palbociclib. J BJAB cells were infected with KSHV and selected with hygromycin (500 µg/mL). Viability of BJAB and BJAB-KSHV was determined in the presence of DON or PALA. K IVIS imaging of NOD-SCID mice engrafted with 2.5 ×106 BCBL-1-luciferase and treated with vehicle control or JHU083. L Survival curve of the 1 ×107 BC-3-luciferase-engrafted NOD-SCID mice treated with vehicle control, JHU083, or Palbociclib. Solid tumors extracted (M) and weighed (N) from the 1 ×107 JSC-1-luciferase-engrafted NOD-SCID mice treated with vehicle control, JHU083, or Palbociclib. Data are presented as mean ± SD of n = 3 biological replicates (7D-7J). Significance was calculated using two-tailed, unpaired Student’s t-test. Source data are provided as a source data file. See also Figs. S7 and S8.