human cdk6 cyclin d3  (Carna Inc)

Journal: Marine Drugs

Article Title: Saccharoquinoline, a Cytotoxic Alkaloidal Meroterpenoid from Marine-Derived Bacterium Saccharomonospora sp.

doi: 10.3390/md17020098

Figure Lengend Snippet: Downregulation of cell cycle checkpoint proteins involved in the G1 phase transition to the S phase. HCT116 cells were treated with 1 (10 μM, +) for 24 hours or 48 hours, and with vehicle (DMSO, 0.1%). The expressions of CDK2, CDK4, CDK6, cyclin D1, and cyclin D3 were illustrated by Western blot analysis. GAPDH was used as the internal control.

Article Snippet: Then, the membrane was incubated with primary antibodies against CDK2, CDK4, CDK6, cyclin D1, cyclin D3, and GAPDH, purchased from Cell Signaling Technology (Danvers, MA, USA), at 4 °C overnight.

Techniques: Sublimation, Western Blot, Control

Journal: Cancer discovery

Article Title: CDK4/6 Inhibition Augments Anti-Tumor Immunity by Enhancing T Cell Activation

doi: 10.1158/2159-8290.CD-17-0915

Figure Lengend Snippet: A, Plot of replicate Z scores from screening of small molecule compounds capable of enhancing IL-2 secretion from stimulated PD-1-Jurkat cells with compounds annotated as GSK3α/β or CDK4/6 inhibitors labeled. B, Quantification of IL-2 levels by ELISA from PD-1-Jurkat cells treated with PMA/ionomycin or 1 µM CDK4/6 inhibitors and stimulated as indicated for 18h. Results shown as mean ± SD (UT, n=2; other conditions, n=5) (*p<0.05). C, Quantification of IL-2 levels by ELISA from primary human CD4+ T cells treated with 100 nM palbociclib or trilaciclib and stimulated as indicated. Results shown as mean ± SD (UT, n=2; other conditions, n=4) (*p<0.05). D, Immunoblot for CDK4 and CDK6 from PD-1-Jurkat cells transiently transfected with the indicated siRNA. E, Quantification of IL-2 levels from PD-1-Jurkat cells after transient transfection with siRNA against Cdk4 or Cdk6 and stimulated as indicated for 18h. Results shown as mean ± SD (n=4) (*p<0.05). Cytokine profiling analysis from human patients using patient-derived organotypic tumor spheroids (PDOTS) cultured in 3-dimensional culturing system at day 1 (F) and day 3 (G). Freshly-obtained patient samples were digested into spheroids and treated with indicated drugs in the 3-D microfluidic system. Cytokine secretion was analyzed by Luminex and expressed as log2-fold change relative to untreated control.

Article Snippet: In vitro Phosphorylation of NFATc3 NMR experiments were performed on a Varian (Agilent DD2 700) spectrometer equipped with a cryogenically cooled probe, and the spectrum was recorded at 287 K. CDK4/cyclin D1 and CDK6/cyclin D3 kinases were purchased from Signalchem.

Techniques: Labeling, Enzyme-linked Immunosorbent Assay, Western Blot, Transfection, Derivative Assay, Cell Culture, Luminex