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. 2021 Oct 6;17(10):e1009971. doi: 10.1371/journal.ppat.1009971

Fig 8. Treatment with LPA and antibody against LPA1 enhances ORFV113 induced p38 phosphorylation.

Fig 8

(A and B) LPA treatment enhances ORFV113 induced p38 phosphorylation. OFTu cells transfected with plasmids pCMV-HA or pORFV113HA for 5 h were induced with LPA harvested at 5, 10 and 20 min. Total cell protein extracts were resolved by SDS-PAGE, blotted and incubated with antibodies against phos-p38, total p38, HA and GAPDH. (B) Densitometry of phos-p38 bands were normalized to total p38 (loading control). Results are average of four independent experiments. Error bars represent mean±SD. Statistical analysis was performed using one-way ANOVA with post hoc Tukey test for multiple comparisons (*, P<0.05; **, P<0.01, NS: non-significant). (C and D) Antibody against LPA1 enhances ORFV113 induced p38 phosphorylation. OFTu cells pretreated with isotype control or LPA1 monoclonal antibody (10 μg/ml) for 45 min were treated with immunoprecipitation products from pCMV-HA or pORFV113HA expressing cells in presence of isotype control or LPA1 monoclonal antibody (10 μg/ml) and harvested at 1 and 3 h. Total cell protein extracts were resolved by SDS-PAGE, blotted and incubated with antibodies against phos-p38 and total p38. (D) Densitometry of phos-p38 bands were normalized as in B. Results are average of five independent experiments. Error bars represent mean ±SD. Statistical analysis was performed as described in B (*,#, P<0.05; **, P<0.01; NS: non-significant).