Syk

Platelets (3 x 10⁸/mL) were preincubated with Y27632 (10 uM), R406 (1 uM), or a combination of Y27632 and R406 for 20 minutes followed by stimulation with oxLDL (50 ug/mL) for 15 seconds and lysis. Samples were then separated by SDS-PAGE and were immunoblotted for phospho-MLCSer19, followed by reprobing for β-tubulin. (Fi) Representative blots. (Fii) Densitometric analysis of 3 independent experiments. *P < .05. Data are presented as mean ± SEM. Experiments were carried out in the presence of apyrase (2 U/mL), indomethacin (10 uM), and EGTA (1 mM).

BCWM.1 and MWCL-1 cells treated with either vehicle or fostamatinib at the indicated concentrations for two hours, then protein extracts were analyzed by Western blotting for the activation status of MEK, p44/42 MAPK, and Akt with phospho-specific antibodies.

(C) Z-138 and JEKO-1 cells were simultaneously  exposed  to sorafenib and R406  at  the  indicated doses, and cell viability was determined at 48 hours by annexin  V/PI  staining.  Bars represent the mean ± SD of 3 independent experiments. CI value is indicated for each combination. (D)  Primary MCL cells from 7 patients were simultaneously exposed to sorafenib and R406 at the indicated doses for 48 hours, and cell viability was determined as above. Bars represent the mean ± SEM of all the samples analyzed. CI value is indicated for each combination.

Ptpn6fl/fl and Inpp5dfl/fl ALL cells carrying 4-OHT-inducible Cre (Cre) or an empty vector were pre-treated with PRT062607 (2.5 µmol/L) for 2 days. Deletion of Ptpn6 (i) or Inpp5d (j) was induced by addition of 4-OHT and relative changes of GFP1 cells were monitored by flow cytometry.

B cells were stimulated by irradiated CD40L+ L cells. Forward scatter vs. BrdU incorporation analysis (day 5) in the gated CD19+ CLL cells (D). The distribution of cells with high FSC (blasts, right quadrants) and proliferation (BrdU+, upper quadrants) are shown.

Influence of piceatannol and resveratrol on hydrogen peroxide production by human subcutaneous fat tissue. H202 release was monitored on a 30-min period without any added agent, except the chromogenic mixture (basal), or with NADPH oxidase inhibitor (apocynin 10 μM, shaded column), each stilbene at 1 mM, or MAO and SSAO inhibitors (pargyline plus semicarbazide at 1 mM: parg + semi, dotted column). Each column is the mean ± SEM of 16 homogenates from obese patients. Different from basal hydrogen peroxide production at: **P < 0.01.

WT mouse primary macrophages were untreated as control (lane 1) or treated with 0.1μg/ml E. coli LPS (lane 5-8), 10μM E-64 (lane 2 and 6), 250μM PMSF (lane 3 and 7), or 20μM MNS (lane 4 and 8). The cells were continuously cultured for 16 hrs. Extracts from whole cells or nuclei were separately purified and subjected to WB analysis with antibody against pERK2, LITAF, or actin/tubulin as control.

(B) SYK(pY352) and PLCγ2(pY759) in CVID patients' CD21^low B cells untreated (filled circles), in the presence of Entospletinib (open circle), or after washout of the inhibitor (gray circles) (n = 5).

Platelets (3 x 10⁸/mL) were preincubated with Y27632 (10 uM), R406 (1 uM), or a combination of Y27632 and R406 for 20 minutes followed by stimulation with oxLDL (50 ug/mL) for 15 seconds and lysis. Samples were then separated by SDS-PAGE and were immunoblotted for phospho-MLCSer19, followed by reprobing for β-tubulin. (Fi) Representative blots. (Fii) Densitometric analysis of 3 independent experiments. *P < .05. Data are presented as mean ± SEM. Experiments were carried out in the presence of apyrase (2 U/mL), indomethacin (10 uM), and EGTA (1 mM).

BCWM.1 and MWCL-1 cells treated with either vehicle or fostamatinib at the indicated concentrations for two hours, then protein extracts were analyzed by Western blotting for the activation status of MEK, p44/42 MAPK, and Akt with phospho-specific antibodies.

(C) Z-138 and JEKO-1 cells were simultaneously  exposed  to sorafenib and R406  at  the  indicated doses, and cell viability was determined at 48 hours by annexin  V/PI  staining.  Bars represent the mean ± SD of 3 independent experiments. CI value is indicated for each combination. (D)  Primary MCL cells from 7 patients were simultaneously exposed to sorafenib and R406 at the indicated doses for 48 hours, and cell viability was determined as above. Bars represent the mean ± SEM of all the samples analyzed. CI value is indicated for each combination.

Ptpn6fl/fl and Inpp5dfl/fl ALL cells carrying 4-OHT-inducible Cre (Cre) or an empty vector were pre-treated with PRT062607 (2.5 µmol/L) for 2 days. Deletion of Ptpn6 (i) or Inpp5d (j) was induced by addition of 4-OHT and relative changes of GFP1 cells were monitored by flow cytometry.

B cells were stimulated by irradiated CD40L+ L cells. Forward scatter vs. BrdU incorporation analysis (day 5) in the gated CD19+ CLL cells (D). The distribution of cells with high FSC (blasts, right quadrants) and proliferation (BrdU+, upper quadrants) are shown.

Influence of piceatannol and resveratrol on hydrogen peroxide production by human subcutaneous fat tissue. H202 release was monitored on a 30-min period without any added agent, except the chromogenic mixture (basal), or with NADPH oxidase inhibitor (apocynin 10 μM, shaded column), each stilbene at 1 mM, or MAO and SSAO inhibitors (pargyline plus semicarbazide at 1 mM: parg + semi, dotted column). Each column is the mean ± SEM of 16 homogenates from obese patients. Different from basal hydrogen peroxide production at: **P < 0.01.

WT mouse primary macrophages were untreated as control (lane 1) or treated with 0.1μg/ml E. coli LPS (lane 5-8), 10μM E-64 (lane 2 and 6), 250μM PMSF (lane 3 and 7), or 20μM MNS (lane 4 and 8). The cells were continuously cultured for 16 hrs. Extracts from whole cells or nuclei were separately purified and subjected to WB analysis with antibody against pERK2, LITAF, or actin/tubulin as control.

(B) SYK(pY352) and PLCγ2(pY759) in CVID patients' CD21^low B cells untreated (filled circles), in the presence of Entospletinib (open circle), or after washout of the inhibitor (gray circles) (n = 5).