Journal: Scientific reports

Article Title: Angiopathic activity of LRG1 is induced by the IL-6/STAT3 pathway.

doi: 10.1038/s41598-022-08516-2

Figure Lengend Snippet: Figure 5. Tocilizumab antibody blocks LRG1 gene expression. HPMEC cells treated with PBS as control (Ctl), 50 ng/ml IL-6, and 20 μg/ml Tocilizumab (Toc) antibody in combination with IL-6 as indicated. RNA and protein were extracted and subjected to RT-qPCR and western blotting. (a) Human LRG1 gene expression normalised to HPRT housekeeping genes. Protein expression of phosphorylated and total STAT3 (b), SMAD1/5 (c) and SMAD2 (d). Quantification was performed by densitometric analysis of normalised phosphorylated to normalised total protein using ImageJ. Representative images of cropped blots (original blots are provided in the supplementary information). N ≥ 3 independent experiments. ANOVA with multiple comparisons test (Tukey’s), Mean ± SEM, * ≤ 0.05, ** ≤ 0.01, *** ≤ 0.001.

Article Snippet: HPMEC were serum-starved overnight and then treated with vehicle (PBS), 50 ng/ml IL-6 alone or in combination with 20 μg/ml anti-IL-6R Tocilizumab (Toc) antibody (Novus Biotechne (NBP2-75192) for 24 or 48 h. RNA and protein were collected, and human LRG1 gene expression was assessed by RT-qPCR.

Techniques: Gene Expression, Control, Quantitative RT-PCR, Western Blot, Expressing

Journal:

Article Title: The Phenotypic Plasticity of Myeloma Plasma Cells as Expressed by Dedifferentiation into an Immature, Resilient, and Apoptosis-Resistant Phenotype

doi: 10.1158/1078-0432.CCR-05-0523

Figure Lengend Snippet: A–B, blocking IL-6 activity did not reverse the immature phenotype of cocultured myeloma cells. CD45/38 flow cytometry of multiple myeloma plasma cells from two patients before culture (precultured), after 6 weeks in culture with osteoclasts (cocultured), and after 4 additional weeks in culture with osteoclasts, anti-IL6, and anti-IL6R. Note an up-regulation of CD45 expression by osteoclasts that was not reversed by inhibition of IL-6 activity.

Article Snippet: Reagents and kits Neutralizing polyclonal antibodies against human interleukin-6 (IL-6) and human IL-6 receptor (IL6R) were obtained from R&D Systems, Inc. (Minneapolis, MN).

Techniques: Blocking Assay, Activity Assay, Flow Cytometry, Clinical Proteomics, Expressing, Inhibition