Additional file 2: Figure S2. of Lysophosphatidic acid via LPA-receptor 5/protein kinase D-dependent pathways induces a motile and pro-inflammatory microglial phenotype

PKD family inhibition abrogates LPA-mediated downstream signaling. (A) BV-2 microglia cells were cultured in 6-well plates, serum-starved overnight and preincubated with CRT0066101 (‘CRT’, 1 μM) for the indicated time periods before incubation with LPA (1 μM) or LPA (1 μM) plus CRT. Cells incubated only with 0.1% BSA or CRT (1 μM) were used as negative control. The phosphorylation states of PKDs, JNK, AKT, ERK1/2, and p38 were detected by immunoblotting and one representative blot for each protein is shown. (B) Densitometric analysis of western blots (N = 3). Results are presented as mean values + SEM (***p < 0.001; compared to control; ## p < 0.01, ### p < 0.001 LPA plus CRT versus LPA; one-way ANOVA with Bonferroni correction). (PPT 777 kb

Additional file 5: Figure S5. of Lysophosphatidic acid via LPA-receptor 5/protein kinase D-dependent pathways induces a motile and pro-inflammatory microglial phenotype

The phosphorylation of pro-inflammatory transcription factors is under PKD family control. BV-2 cells, serum-starved overnight and treated with LPA (1 μM) or LPA (1 μM) in the presence of (A) CRT0066101 (1 μM) for the indicated time periods. Cells incubated only with 0.1% BSA or CRT (1 μM) were used as negative control. The phosphorylation state of p65-NF-κB, STAT1, STAT3, and c-Jun was detected by western blotting. One representative blot is shown. (B) Densitometric analysis of western blots (N = 3). Results are presented as mean values + SEM (**p < 0.01, ***p < 0.001 compared to control; ## p < 0.01, ### p < 0.001 LPA plus CRT versus LPA; one-way ANOVA with Bonferroni correction). (PPT 500 kb

Additional file 6: Figure S6. of Lysophosphatidic acid via LPA-receptor 5/protein kinase D-dependent pathways induces a motile and pro-inflammatory microglial phenotype

PKD family members control the secretion of pro-inflammatory cytokines and chemokines. BV-2 cells were cultured on 12-well plates; serum-starved o/n and the supernatants were collected after incubation with DMSO, DMSO plus LPA (1 μM) or LPA (1 μM) plus CRT (1 μM). ELISAs were used to quantitate IL-6, IL-1β, CXCL10 (IP-10), TNF-α, CXCL2 (MIP-2), and CCL5 (RANTES) concentrations. Results shown represent mean + SD from three independent experiments performed in triplicate (*p < 0.05; **p < 0.01 compared to vehicle control; # p < 0.05, ## p < 0.01; CRT + LPA compared to LPA treated cells; one-way ANOVA with Bonferroni correction). (PPT 174 kb

Additional file 1: of Overexpression of KCNJ3 gene splice variants affects vital parameters of the malignant breast cancer cell line MCF-7 in an opposing manner

Supplementary Figures. (PDF 1221 kb