Figure 5. Nuclear p38MAPK localization is necessary for HCF migration. HCFs were seeded on collagen in SSFM at 1×10⁵ cells/ml in a 24 well dish. The next day cells were scratch-wounded and incubated in (A, F, K) p38MAPK inhibitor, 10 μM SB202190, (B, G, L) DMSO, (C, H, M) TGFβ1 antibody, (D, I, N) Control IgG, or (E, J, O) TGFβ RI (ALK5) inhibitor 10 μM SB431542. After 4 h cells were fixed and immunostained for p38MAPK (A-E) or SMAD 2/3 (F-J). Arrows denote the nuclei of leading edge cells in which p38MAPK and SMAD 2/3 were either localized or excluded (Bar=50 μm) or after 18 h cells were imaged (K-O). Bar=200 μm. DMSO is the control for addition of SB202190 or SB431542. Quantification of all data are shown in the bar graphs below the images. Left to right: Exclusion of p38MAPK from the nucleus in leading edge cells, exclusion of SMAD 2/3 from the nucleus in leading edge cells, cell migration into the wound. Nuclear localization was counted using Image J software. Two non-biased people scored cell migration, 0 (less migration) to 5 (most migration). **p-value <0.01, ***p-value <0.001. NS=not significant. Experiments were repeated at least three times with similar results.