Fig 2: Infusion of LIMKi 3 into the medial prefrontal cortex (mPFC) produced antidepressant-like effects in the chronic unpredictable mild stress (CUMS) model of depression. (A) Schematic timeline of the experimental procedures. (B) LIMKi 3 fully prevented the CUMS-induced depressive-like behaviors in mice, as revealed by the forced swim test (FST), tail suspension test (TST), and sucrose preference test (n = 10). (C) Both the (CUMS + 100 µM LIMKi 3)-treated and (CUMS + 200 µM LIMKi 3)-treated mice had significantly lower levels of pLIMK1, pLIMK2, and pCofilin in the mPFC than the CUMS-treated mice (n = 6). The expression of total LIMK1, LIMK2, and Cofilin in the mPFC remain unchanged among all groups (n = 6). All results are expressed as the means ± SEM; *P < .05, **P < .01; n.s., no significance. The comparisons were made by 2-way ANOVA followed by Bonferroni’s test.

Fig 3: Infusion of LIMKi 3 into the medial prefrontal cortex (mPFC) produced antidepressant-like effects in the chronic restraint stress (CRS) model of depression. (A) Schematic timeline of the experimental procedures. (B) LIMKi 3 significantly attenuated the CRS-induced depressive-like behaviors in mice, as revealed by the forced swim test (FST), tail suspension test (TST), and sucrose preference test (n = 10). (C) Both the (CRS + 100 µM LIMKi 3)-treated and (CRS + 200 µM LIMKi 3)-treated mice had significantly lower levels of pLIMK1, pLIMK2, and pCofilin in the mPFC than the CRS-treated mice (n = 6). The expression of total LIMK1, LIMK2, and Cofilin in the mPFC remained unchanged among all groups (n = 6). All results are expressed as the means ± SEM; **P < .01; n.s., no significance. The comparisons were made by 2-way ANOVA followed by Bonferroni’s test.

Fig 4: Effects of KIF and AUY-922 on lung endothelial cells.Western Blot analysis of (A) phosphorylated Cofilin (pCofilin) and Cofilin (B) phosphorylated MLC2 (pMLC2) and MLC2 after 24 h treatment of BPAEC with either KIF (5 µM) or vehicle (0.1% DMSO) and post-treatment with AUY-922 (2 µM) or vehicle (0.1% DMSO) for 16 h. The blots shown are representative of 4 independent experiments. The signal intensity of the bands was analyzed by densitometry. Protein levels of pCofilin and pMLC2 were normalized to Cofilin and MLC2 respectively. *P < 0.05, **P < 0.01 vs vehicle. Means ± SEM. Western Blot analysis of (C) phosphorylated MLC2 (pMLC2) and MLC2 after 24 h treatment of HuLEC with either KIF (5 µM) or vehicle (0.1% DMSO); and post-treatment with AUY-922 (2 µM) or vehicle (0.1% DMSO) for 16 h. The blots shown are representative of 3 independent experiments. The signal intensity of the pMLC2 was analyzed by densitometry. Protein levels of pMLC2 were normalized to MLC2. *P < 0.05 vs vehicle. Means ± SEM. (D) Confluent monolayers of BPAEC were pre-treated with either vehicle (0.1% DMSO) or KIF (25 µM) (red arrow) for 18 h, followed by treatment with either vehicle (0.1% DMSO) or 5 µM of AUY-922 (green arrow). A gradual increase in endothelial permeability (reduced TEER) was observed in KIF treated cells (red line). AUY-922 significantly reduced the endothelial permeability (increased TEER) in both KIF-pretreated (blue line) and vehicle-treated cells (green line). (E) Cells were incubated with either VEH (0.1% DMSO) or KIF (0.01, 0.1, 1, 10, 100, 200 µM) or AUY-922 (0.01, 0.1, 1, 10, 100, 200 µM) for 24 h. Cellular viability was evaluated by employing the MTT assay. ***P < 0.001 vs VEH, n = 3. Means ± SEM. (for interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Fig 5: Lack of P53 potentiates LPS-triggered lung inflammatory responses in vivo. Wild type and P53 KO mice were treated with vehicle (saline) or LPS (1.6 mg/kg) via an intratracheal injection. Measurements of total protein levels in the BALF of wild type (2A) and P53 KO mice (2B). **P ?< ?0.01, ***P ?< ?0.001 vs vehicle (VEH), n ?= ?4, Means ?± ?SEM. Western Blot analysis of (2C) pCofilin and Cofilin in lung homogenates of wild type mice. The signal intensity of the bands was analyzed by densitometry. Protein levels of pCofilin were normalized to Cofilin. *P ?< ?0.05 vs vehicle (VEH), n ?= ?4. Means ?± ?SEM. Western Blot analysis of (2D) pCofilin and Cofilin in lung homogenates of P53 KO mice. The signal intensity of the bands was analyzed by densitometry. Protein levels of pCofilin were normalized to Cofilin. *P ?< ?0.05 vs vehicle (VEH), n ?= ?4. Means ?± ?SEM.