phosphatase

Effect of FK506 on cytosolic calcium homeostasis induced by TG and thrombin in human platelets. (A-B) Human platelets were suspended in a Ca 2+-free medium (100 µM of EGTA was added as indicated by arrowhead), and preincubated for 5 min at 37oC in the absence (solid black traces) or presence of increased concentrations (0.01-100 µM) of FK506 (light-doted and dark solid-grey traces, respectively). Cells were then stimulated with TG (200 nM; A) or Thr (0.1 U/mL; B) and 3 min later 300 μM of CaCl2 was added to extracellular medium to visualize calcium entry.

Repression of BSEP gene expression upon treatment with reported BSEP inhibitors in human primary hepatocytes. BSEP mRNA expression was measured using real-time PCR in hepatocytes from at least three donors upon treatment with BSEP inhibitors as described under Materials and Methods. All the data are expressed as mean ?S.D. (n = 3).

(A-B)Twenty-four hours after plating 1×105 shh-EGFP cells, media was changed to serum-free media containing vehicle (Vh), DMSO, CCT007093 (CCT, 5µM), and/or Shh (3µg/mL). Cells were also transduced with LentiORF-RFP-CTRL or LentiORF-YFP-WIP1 lentivirus. 48 hours later, cells were fixed in 4% paraformaldehyde, permeabilized, incubated with α-WIP1 or α-Ki-67 antibody, and mounted using media containing DAPI (left-hand panels). Quantification of cells, per high power field (HPF), that express green fluorescent protein (GFP) or Ki-67 (right-hand panels). Bars, mean of cell counts from 10 representative fields for each experimental condition.

LB-100 induced HDAC2 S394 phosphorylation.

BMDMs and BMDCs were pretreated with NSC87877 (1 μM) for 1 h followed by stimulation with anti-CD180 Ab (0.2 μg/ml). Then the cells were stimulated with R837 (1 μg/ml) and CpG 1826 (0.5 μM). ELISA analysis of the levels of IL-6 and IL-12 secreted by BMDMs (E) and BMDCs (F) at 24 h.

Effect of FK506 on cytosolic calcium homeostasis induced by TG and thrombin in human platelets. (A-B) Human platelets were suspended in a Ca 2+-free medium (100 µM of EGTA was added as indicated by arrowhead), and preincubated for 5 min at 37oC in the absence (solid black traces) or presence of increased concentrations (0.01-100 µM) of FK506 (light-doted and dark solid-grey traces, respectively). Cells were then stimulated with TG (200 nM; A) or Thr (0.1 U/mL; B) and 3 min later 300 μM of CaCl2 was added to extracellular medium to visualize calcium entry.

Repression of BSEP gene expression upon treatment with reported BSEP inhibitors in human primary hepatocytes. BSEP mRNA expression was measured using real-time PCR in hepatocytes from at least three donors upon treatment with BSEP inhibitors as described under Materials and Methods. All the data are expressed as mean ?S.D. (n = 3).

(A-B)Twenty-four hours after plating 1×105 shh-EGFP cells, media was changed to serum-free media containing vehicle (Vh), DMSO, CCT007093 (CCT, 5µM), and/or Shh (3µg/mL). Cells were also transduced with LentiORF-RFP-CTRL or LentiORF-YFP-WIP1 lentivirus. 48 hours later, cells were fixed in 4% paraformaldehyde, permeabilized, incubated with α-WIP1 or α-Ki-67 antibody, and mounted using media containing DAPI (left-hand panels). Quantification of cells, per high power field (HPF), that express green fluorescent protein (GFP) or Ki-67 (right-hand panels). Bars, mean of cell counts from 10 representative fields for each experimental condition.

LB-100 induced HDAC2 S394 phosphorylation.

BMDMs and BMDCs were pretreated with NSC87877 (1 μM) for 1 h followed by stimulation with anti-CD180 Ab (0.2 μg/ml). Then the cells were stimulated with R837 (1 μg/ml) and CpG 1826 (0.5 μM). ELISA analysis of the levels of IL-6 and IL-12 secreted by BMDMs (E) and BMDCs (F) at 24 h.