Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Systemic lupus erythematosus immune complexes increase the expression of SLAM family members CD319 (CRACC) and CD229 (LY-9) on plasmacytoid dendritic cells and CD319 on CD56(dim) NK cells.

doi: 10.4049/jimmunol.1301022

Figure Lengend Snippet: FIGURE 5. pDCs lack the activating adaptor molecules but express inhibitory adaptor molecules of SLAM receptors. (A) IFN-a production by RNA-IC–stimulated pDCs after cross-linking of CD319, CD229, or BDCA-2 by plate-bound mAbs. IFN-a levels were determined in 20-h cell culture supernatants by an immunoassay and normalized to the IFN-a production in cultures without plate-bound mAb. The dashed line indicates 100%. Data (mean + SD) are from three donors from two independent experiments. Expression of EAT2 and SAP (B) and SHIP-1, SHP-1, SHP- 2, and CSK (C) in cell lysates from isolated pDCs or NK cells from two healthy donors was determined by Western blot. Expression of b-actin was used as control.

Article Snippet: Rabbit polyclonal Abs to Ewing’s sarcoma-activated transcript 2 (EAT2), SHIP-1, SHP-2, CSK (Proteintech Group, Chicago, IL), and a rabbit mAb to SHP-1 (EPR5519; Epitomics, Burlingame, CA), followed by HRP-conjugated goat anti-rabbit IgG (H+L) (Invitrogen), were used to detect the indicated signaling molecules.

Techniques: Cell Culture, Expressing, Isolation, Western Blot, Control

Journal: Oncogenesis

Article Title: A functional CRISPR/Cas9 screen identifies kinases that modulate FGFR inhibitor response in gastric cancer

doi: 10.1038/s41389-019-0145-z

Figure Lengend Snippet: a Western blot confirmed that ILK and CSK gene expression was effectively knocked down in KatoIII cells. Cells were transfected with 10 nmol/l oligos and were harvested 72 h. ILK (#3856) and CSK (#4980) antibodies were obtained from Cell Signaling. Vinculin (#13901, Cell Signaling) was the loading control. b Upon AZD4547 treatment, knocking down ILK decreased cell viability while knocking down CSK increased cell viability. c Knocking down ILK or CSK alone does not affect KatoIII cell viability. KatoIII cells were seeded in 6-well plates at 300,000 cells/well the day before transfection and were transfected with 10 nM siRNA oligos using LipoJet (SignaGen Laboratories). The Dicer-substrate short interfering RNAs (DsiRNAs) targeting CSK (CSK_1 Reference #146599035 and CSK_2 #146599032), ILK (ILK_1 Reference #146599024 and ILK_2 #146599021) as well negative control oligos (NTC) were obtained from IDT. The viability was measured 6 days by CellTiter-Glo after treated with AZD4547 or plain median. Representative data from one of three independent experiments are shown. The data are measured in hextuple as mean ± s.e.m. **** P < 0.0001; ** P < 0.01. The P values were calculated by Dunnett’s multiple comparisons test

Article Snippet: Cells were transfected with 10 nmol/l oligos and were harvested 72 h. ILK (#3856) and CSK (#4980) antibodies were obtained from Cell Signaling.

Techniques: Western Blot, Gene Expression, Transfection, Control, Negative Control