Fig. 1

REG inhibits CSF1/CSF1R signaling in macrophages in vitro. A Schematic diagram of CSF1R signaling relating to this study. B Western blot analysis of total lysates (40 µg/lane) from mouse RAW264.7 cells treated as indicated. Tyrosine phosphorylation sites correspond to the murine CSF1R. The two phosphorylation sites were analyzed on separate blots together with the total CSF1R and β-actin as loading control. The pY721 blot was re-probed for total/pAKT and total/pERK was analyzed on a new blot. Fluorescent signals were captured densitometrically and used to determine IC₅₀. The whole blot figure is shown in Fig. S3 (see Additional file 2). C Example IC₅₀ determination using GraphPad software. DMSO without and with CSF1 stimulation was used as controls. D Cellular IC₅₀ values of various kinases are given as means with standard deviation. The number of assay replicates is given in brackets. *IC₅₀ for pCSF1R is derived directly from the densitometric values of the respective pCSF1R bands without relating to the signal of total CSF1R, which was destabilized by M-4 and M-5 (see Additional file 2: Fig. S2). ^#Data for M-2 and M-5 taken from Zopf et al., 2016 [5]. IL, interleukin; K, competitive kinase binding assay; WB, western blot.