Journal: Frontiers in Immunology

Article Title: TLQP-21 is a low potency partial C3aR activator on human primary macrophages

doi: 10.3389/fimmu.2023.1086673

Figure Lengend Snippet: hTLQP-2 and mTLQP-21 activate ERK signalling in CHO-K1 and CHO-C3aR cells. hTLQP-21, mTLQP-21 and plasma-derived human C3a were tested in (A) non-transfected CHO-K1 or (B) CHO cells stably expressing human C3aR. CHO cells were serum-starved overnight and then stimulated with various ligands for 10 min before being lysed. The phospho-ERK1/2 content in the lysate was measured and expressed as fold-baseline before being combined. The maximum relative pERK1/2 activity induced by each ligand is shown in (C) . Data represent mean ± S.E.M. of triplicate measurements from 3-4 independent experiments (n = 3-4). Two-way ANOVA with Dunnett’s post hoc analysis. * P < 0.05, *** P < 0.001, **** P < 0.0001. Ligand treated versus medium treated cells for each cell line.

Article Snippet: Non-transfected CHO-K1 cells or CHO cells stably expressing the human C3aR (CHO-C3aR; Product # ES-730-C, PerkinElmer, Melbourne, Australia) were maintained in Ham’s F12 medium containing 10% foetal bovine serum (FBS), 100 IU/ml penicillin and 100 μg/ml streptomycin, with additional 400 μg/ml G418 ( In vivo Gen, San Diego, USA) added for CHO-C3aR cell culture.

Techniques: Clinical Proteomics, Derivative Assay, Transfection, Stable Transfection, Expressing, Activity Assay

Journal: Frontiers in Immunology

Article Title: TLQP-21 is a low potency partial C3aR activator on human primary macrophages

doi: 10.3389/fimmu.2023.1086673

Figure Lengend Snippet: Summary of potencies and activities of TLQP-21 tested on CHO-C3aR, HMDM and BMDM.

Article Snippet: Non-transfected CHO-K1 cells or CHO cells stably expressing the human C3aR (CHO-C3aR; Product # ES-730-C, PerkinElmer, Melbourne, Australia) were maintained in Ham’s F12 medium containing 10% foetal bovine serum (FBS), 100 IU/ml penicillin and 100 μg/ml streptomycin, with additional 400 μg/ml G418 ( In vivo Gen, San Diego, USA) added for CHO-C3aR cell culture.

Techniques: Activity Assay

Journal: Frontiers in Immunology

Article Title: TLQP-21 is a low potency partial C3aR activator on human primary macrophages

doi: 10.3389/fimmu.2023.1086673

Figure Lengend Snippet: TLQP-21 triggers ERK signalling through C3aR in murine bone marrow-derive macrophages. BMDMs (90,000/well) from wildtype mice (A) or with C3aR knockout mice (B) , were serum-starved overnight and then stimulated with respective ligands at the indicated concentrations for 10 min. The phospho-ERK1/2 content in the cell lysate was measured and normalised to the medium only-treated levels before being combined. Data represent mean ± S.E.M. of triplicate measurements using cells from 3 mice (n = 3).

Article Snippet: Non-transfected CHO-K1 cells or CHO cells stably expressing the human C3aR (CHO-C3aR; Product # ES-730-C, PerkinElmer, Melbourne, Australia) were maintained in Ham’s F12 medium containing 10% foetal bovine serum (FBS), 100 IU/ml penicillin and 100 μg/ml streptomycin, with additional 400 μg/ml G418 ( In vivo Gen, San Diego, USA) added for CHO-C3aR cell culture.

Techniques: Knock-Out