Fig 2: MARCKS is involved in CAF proliferation, chemotherapeutical resistance and migrationA. Western blot analysis of MARCKS and p-MARCKS in purified CAFs from EOC tumor tissues obtained either at initial diagnosis or at disease recurrence after several cycles of chemotherapeutical intervention. B. Immunoblotting of MARCKS and p-MARCKS protein expression in MRC5-CAF cells in the absence or presence of si-NC or si-MARCKS or enzastaurin (Enza, 1 µM) treatment for 48 h. C. CCK8 assay detecting relative cell numbers of MRC5-CAFs in the si-NC or si-MARCKS or Enza (1 µM) intervention groups for various duration (1-5 days). D. Detection of relative cell viability in the si-NC or si-MARCKS or Enza (1 µM) pretreated groups of MRC5-CAFs, in the absence or presence of cDDP or taxol intervention at various dosage for 48 h. E. Representative images demonstrating and statistical analysis of wound closure capacity of MRC5-CAFs in si-NC or si-MARCKS or Enza (1 µM) treatment groups at 0 h and 12 h time points. F. Spearman correlation analysis of MARCKS expression with that of normalized EMT score in integrated large cohorts of ovarian cancer [38]. G. Western blot analysis of PI3K, p-PI3K, AKT and p-AKT expression in MRC5-CAFs and primary ovarian CAFs, in the presence of si-NC or si-MARCKS or Enza (1 µM) treatment for 72 h. GAPDH served as the loading control. Data are expressed as mean ± s.e.m. (*P < 0.05; **P < 0.01).

Fig 3: DHA eliminates apoptotic cell death and restores AKT phosphorylation in primary cultured Schwann cells (pSC) under PA-LTx. pSC were treated with BSA alone (CTL), with PA:BSA (2:1) alone, with DHA (50 µM) alone or co-treated with PA:BSA (2:1) and DHA (50 µM) for 48 hr. Nuclear morphology was determined by Hoechst staining. Nuclear condensations are indicated with white arrows (a). The pSC culture lysates were prepared and subjected to Western blot analysis using specific antibodies against AKTp-Ser473, AKTp-Thr308, and total AKT protein. The blots were then analyzed using the Li-Cor Odyssey system. A representative Western blot is shown by each bar graph (b). The data represent mean ± SEM of at least four independent experiments. *p < 0.05 are shown above the bars when compared to control groups. # p < 0.05 and ## p < 0.01 are shown above lines when compared between two linked groups

Fig 4: The protective effect of DHA on primary cultured Schwann cells (pSC) under PA-LTx is abolished by PI3K/AKT inhibitors. pSC cultures were treated with DHA (50 µM) in the presence or absence of PI3K inhibitors LY290042 (40 µM) or BKM120 (2 µM) (a). The pSC cultures were treated with PA:BSA (2:1), PA + DHA in a presence or absence of PI3K inhibitors LY290042 (40 µM), or BKM120 (2 µM) (b). Cell viability was measured at 48 hr by crystal violet assay. Cells morphology was visualized at 48 hr by Hoffman modulation contrast microscopy. Membrane blebs and cell shrinkage are indicated with white arrows (c). The data represent mean ± SEM of at least five independent experiments **p < 0.05, ***p < 0.001 when compared between two linked groups

Fig 5: ICA ? restores SRPK1/Akt/eNOS pathway in PA-treated HUVECs. HUVECs were treated with 5 µM SPHINX31, followed by stimulation with PA (150 µM) in the presence or absence of ICA ? (5 µM) for 24 h that was pretreated for 1 h. The levels of eNOS, phosphor-eNOS (Ser1177), Akt, phosphor-Akt (Ser473), and SRPK1 were determined by western blot (A) with the quantifications of the related proteins (B). The immunofluorescence images of the NO probe were observed using 40 × fluorescence microscopy (C) with the quantifications of mean fluorescence intensity (D). The data are presented as the mean ± SD of three independent experiments. Significance was determined by one-way ANOVA with a least significant difference post hoc test. ns, p > 0.05; *, p < 0.05 vs Ctrl.