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: bioRxiv

Article Title: The c-Src inhibitor eCF506 diminishes opioid tolerance creating bias against β-arrestin2 recruitment

doi: 10.1101/2025.07.01.662507

Figure Lengend Snippet: Concentration-response relationships of DAMGO as a recruiter of β-arrestin2 to μ receptors in PathHunter CHO cells transiently overexpressing ( A ) full-length (residues 1-536; WT) versus C-terminally truncated (residues 1-249) human c-Src and ( B ) functional (residues 250-536) versus catalytically non-functional (residues 250-536(K298M)) N-terminally truncated human c-Src. The insert in (A) demonstrates overexpression of C-terminally truncated c-Src using an antibody against HA (1:1000). Overexpression of other c-Src mutants is depicted in Supplementary Figure 5A. WT responses in (B) are depicted as a dashed line. Data are expressed as a percentage of maximum luminescence produced by DAMGO in cells overexpressing WT c-Src (% WT). Data from individual replicates were plotted and fitted with logistics functions to derive efficacy (E MAX ) and potency (EC 50 ) parameters, presented in . Data in A and B are the mean ± SEM of 9 replicates.

Article Snippet: All constructs are based on human c-Src WT (1-536) c-Src (plasmid #42202; RRID: Addgene_42202), c-terminus truncated (1-249) c-Src (plasmid #42204; RRID: Addgene_42204), n-terminus truncated (250-536) c-Src (plasmid #42208; RRID: Addgene_42208), and kinase dead (K298M) c-Src (plasmid #42210; RRID: Addgene_42210), were donated by Dr Robert Lefkowitz.

Techniques: Concentration Assay, Functional Assay, Over Expression, Produced

Journal: bioRxiv

Article Title: The c-Src inhibitor eCF506 diminishes opioid tolerance creating bias against β-arrestin2 recruitment

doi: 10.1101/2025.07.01.662507

Figure Lengend Snippet: Concentration-response relationships of DAMGO to recruit β-arrestin2 to human μ receptors in PathHunter CHO cells transiently overexpressing ( A ) full-length (residues 1-536; WT), ( B ) C-terminally truncated (residues 1-249) or ( C ) N-terminally truncated and catalytically inactive (residues 250-536(K298M)) human c-Src following 16-h exposure to PP2, PP3, eCF506 (300 nM) or an equal volume of DMSO. Data are expressed as a percentage of maximum luminescence (% Max RLU) produced by DAMGO in DMSO exposed cells present in duplicate on each plate i.e., cells overexpressing (A) WT, (B) Src 1-249 or (C) Src 250-536(K298M) are each expressed relative to the maximum β-arrestin2 recruitment of each construct caused by DAMGO. Data from individual replicates were plotted and fitted with logistics functions to derive efficacy (E MAX ) and potency (EC 50 ) parameters, presented in Supplementary Table 3. Data are the mean ± SEM of 5-6 replicates.

Article Snippet: All constructs are based on human c-Src WT (1-536) c-Src (plasmid #42202; RRID: Addgene_42202), c-terminus truncated (1-249) c-Src (plasmid #42204; RRID: Addgene_42204), n-terminus truncated (250-536) c-Src (plasmid #42208; RRID: Addgene_42208), and kinase dead (K298M) c-Src (plasmid #42210; RRID: Addgene_42210), were donated by Dr Robert Lefkowitz.

Techniques: Concentration Assay, Produced, Construct