Journal: Cells

Article Title: Enhanced Migration of Fuchs Corneal Endothelial Cells by Rho Kinase Inhibition: A Novel Ex Vivo Descemet’s Stripping Only Model

doi: 10.3390/cells13141218

Figure Lengend Snippet: Activation of Rac1 driving lamellipodia formation at the leading edge enhancing cell migration. Rac1 activation in normal and FECD cells at ( A ) baseline and ( B ) after the treatment with ripasudil compared to baseline. ( C ) The expression of ARPC2 at the leading edge in normal and FECD cells with and without ripasudil. Pearson’s correlation between normal and FECD cells at ( D ) baseline and ( E ) after the treatment with ripasudil compared to baseline. * p < 0.05; ** p < 0.01; *** p < 0.001. The data are expressed as mean ± SEM. Blue: nucleus; Red: actin; Green: ARPC2; Yellow: merging of actin and ARPC2. Scale: 10 µm.

Article Snippet: All of the cell lines were plated on a 96-well plate, treated with 1 μM of ripasudil or left untreated as controls, and incubated for 24 h before the GTPase assay was performed using the GTPase-Glo Assay kit (Promega, Madison, WI, USA) and Active Rac1 kit (SignalChem, Richmond, BC, Canada).

Techniques: Activation Assay, Migration, Expressing

Journal: Oncotarget

Article Title: Oncogenic features of neuromedin U in breast cancer are associated with NMUR2 expression involving crosstalk with members of the WNT signaling pathway.

doi: 10.18632/oncotarget.16121

Figure Lengend Snippet: Figure 8: NMU signaling modulates the WNT receptor pathway in NMUR2-positive SKBR3 breast cancer cells. (A) Real-time PCR-based validation of candidate NMU downstream genes in independent stably transfected SKBR3 NMU (n=5) and mock clones (n=5). * P < 0.05; ns: not significant (Mann-Whitney-U test). (B) Representative western blots showing differential protein expression of candidate NMU downstream genes in independent stably transfected SKBR3 NMU and mock clones. Loading controls: β-actin, β-tubulin, total RAC1. All experiments were performed in triplicate. (C) Densitometrical evaluation of the western blot results shown in B depicted as box plots. Box plot showing RAC1-GTP in relation to total RAC1 amounts in SKBR3 NMU (n=4) and mock clones (n=4), combines data of three independent experiments. * P < 0.05; ns: not significant (Mann-Whitney-U test). (D) Hypothetical model of NMU’s oncogenic role in dependency of NMUR2 in breast cancer: crosstalk of NMU signaling with WNT, TGFβ and ERK cascade results in decreased expression of the canonical WNT target MYC and enhanced activation of the non-canonical WNT/planar cell polarity (PCP) pathway effector RAC1 among others, contributing to growth inhibition and promotion of cell migration.

Article Snippet: Measurement of RAC1 activation in stable SKBR3 NMU and mock clones was achieved by using the Active RAC1 Detection Kit (#8815, Cell Signaling, Danvers, MA, USA) according to the manufacturer’s instructions.

Techniques: Real-time Polymerase Chain Reaction, Biomarker Discovery, Stable Transfection, Transfection, Clone Assay, MANN-WHITNEY, Western Blot, Expressing, Activation Assay, Inhibition, Migration

Journal: Experimental neurology

Article Title: RvD1binding with FPR2 attenuates inflammation via Rac1/NOX2 pathway after neonatal hypoxic-ischemic injury in rats.

doi: 10.1016/j.expneurol.2019.112982

Figure Lengend Snippet: Fig. 5. Effects of FPR2 siRNA and Rac1 activation CRISPR on infarct area and geotaxis reflex at 24 h post hypoxia-ischemia (HI). (A and B)The infarction area was significantly enlarged with FPR2 siRNA/Rac1 activation CRISPR interventions when compared with respective controls. (C) Interventions with FPR2 siRNA or Rac1 activation CRISPR significantly worsened geotaxis reflex compared with respective controls. ANOVA followed by Tukey test was used for analysis. Data were shown as mean ± SD (n = 6 per group). * P < 0.05 vs. sham group; # P < 0.05 vs. HI + vehicle group, & P < 0.05 vs. HI + RvD1 + scramble siRNA group, @ P < 0.05 vs. control CRISPR. RvD1, resolvin D1.

Article Snippet: 1 μg of Rac1 CRISPR activation plasmid (Santa Cruz, USA) or control CRISPR Activation Plasmid (Santa Cruz, USA) were given via ICV injection as described above at 48 h before HI.

Techniques: Activation Assay, CRISPR, Control

Journal: Experimental neurology

Article Title: RvD1binding with FPR2 attenuates inflammation via Rac1/NOX2 pathway after neonatal hypoxic-ischemic injury in rats.

doi: 10.1016/j.expneurol.2019.112982

Figure Lengend Snippet: Fig. 6. Knockdown of FPR2 reduced the anti-inflammation effect of RvD1 at 24 h post hypoxia-ischemia (HI). (A) Representative Western blot bands. (B–G) Quantification of Western blot bands showed that RvD1 significantly increased the expression of FPR2 (B), whereas decreased the expression of GTP-Rac1 (C), total Rac1(D), NOX2 (E), IL-1β (F), and TNF-α (G) compared with HI + vehicle group. However, FPR2 siRNA reversed the effect of RvD1 compared with HI + RvD1 + scramble siRNA group. ANOVA followed by Tukey test was used for analysis. Data were shown as mean ± SD (n = 6 per group). * P < 0.05 vs. sham group; # P < 0.05 vs. HI + vehicle group; & P < 0.05 vs. HI + RvD1 + scramble siRNA group. RvD1, resolvin D1.

Article Snippet: 1 μg of Rac1 CRISPR activation plasmid (Santa Cruz, USA) or control CRISPR Activation Plasmid (Santa Cruz, USA) were given via ICV injection as described above at 48 h before HI.

Techniques: Knockdown, Western Blot, Expressing

Journal: Experimental neurology

Article Title: RvD1binding with FPR2 attenuates inflammation via Rac1/NOX2 pathway after neonatal hypoxic-ischemic injury in rats.

doi: 10.1016/j.expneurol.2019.112982

Figure Lengend Snippet: Fig. 7. Activation of Rac1 decreased the anti-inflammation function of RvD1 at 24 h post hypoxia-ischemia (HI). (A) Representative Western blot bands. (B–G) Quantification of Western blot bands showed that Rac1 activation CRISPR inhibited the effect of RvD1 on decreasing the expression of GTP-Rac1 (C), total Rac1 (D), NOX2 (E), IL-1β (F), and TNF-α (G) compared with HI + RvD1 + control CRISPR. ANOVA followed by Tukey test was used for analysis. Data were shown as mean ± SD (n = 6 per group). * P < 0.05 vs. sham group; # P < 0.05 vs. HI + vehicle group; & P < 0.05 vs. HI + RvD1 + control CRISPR group. RvD1, resolvin D1.

Article Snippet: 1 μg of Rac1 CRISPR activation plasmid (Santa Cruz, USA) or control CRISPR Activation Plasmid (Santa Cruz, USA) were given via ICV injection as described above at 48 h before HI.

Techniques: Activation Assay, Western Blot, CRISPR, Expressing, Control