Journal: Cell Death & Disease

Article Title: RhoA balances microglial reactivity and survival during neuroinflammation

doi: 10.1038/s41419-023-06217-w

Figure Lengend Snippet: A Primary cortical microglia expressing the Raichu-RhoA biosensor exposed to 1 µg/ml LPS ( n = 6 cells pooled across 3 independent cultures). Panels represent time-lapse FRET/CFP images coded according to the pseudocolor ramp. Graph (mean and SD) displays FRET/CFP ratio changes normalized at 0 min. * p < 0.05 (One-way ANOVA). B Primary cortical microglia expressing the Raichu-RhoA biosensor were exposed to different concentrations of LPS for 20 min ( n = 10 cells pooled across 5 independent cultures). Graph (mean and SD) displays FRET/CFP ratio changes. The blue line shows untreated cells’ mean FRET/Donor ratio changes. The dashed line indicates the IC 50 . C , Primary cortical microglia treated and non-treated (CT) with LPS (1 µg/ml) for 1 h ( n = 10 independent cultures) and immunostained for GTP-RhoA. Graph (mean and SD) shows amounts of GTP-RhoA as mean fluorescent intensity (MFI). **** p < 0.0001 (Mann-Whitney test). D RhoA pull-down on lysates from primary cortical microglia treated and non-treated (CT) with LPS (1 µg/ml) for 1 h ( n = 3 independent cultures). Graph displays mean with SD. * *p < 0.01 (Mann-Whitney test). E HMC3 microglia expressing Raichu-RhoA biosensor treated and non-treated (CT) with 1 µg/ml LPS for 1 h ( n = 100 cells per group from 10 independent experiments). Pseudocolor ramp represents min/max FRET/CFP ratios. Graph (mean and SD) displays FRET/CFP ratio changes. **** p < 0.0001 (unpaired t-test). F RhoA pull-down on lysates from HCM3 microglia treated and non-treated (CT) with LPS (1 µg/ml) for 1 h ( n = 3 independent cultures). Graph displays mean with SD. * p < 0.05 (Mann-Whitney test). G Immunofluorescence images of GTP-RhoA and Iba-1 on cortical tissue sections from brain cortex of adult mice injected with saline or LPS (4 mg/Kg; 24 h). Graph (mean and SD) shows amounts of GTP-RhoA in Iba-1+ cells ( n = 3 mice per group). * p < 0.001 (paired t-test). Scale bars: 10 µm ( A, C , and G ); 20 µm ( E ).

Article Snippet: GTP-RhoA (NewEast Biosciences Cat# 26904, RRID:AB_1961799), RhoA (Abcam Cat# ab68826, RRID:AB_1142593), RhoA (Cell Signaling Technology Cat# 2117, RRID:AB_10693922), Iba-1 (FUJIFILM Wako Shibayagi Cat# 27030, RRID:AB_2314667), phosphorylated Src family (Cell Signaling Technology Cat# 6943, RRID:AB_10013641), Src (Cell Signaling Technology Cat# 2102, RRID:AB_331358), phosphorylated ERK 1/2 (Cell Signaling Technology Cat# 9101, RRID:AB_331646), ERK 1/2 (Cell Signaling Technology Cat# 9102, RRID:AB_330744).

Techniques: Expressing, MANN-WHITNEY, Immunofluorescence, Injection, Saline

Journal: International Journal of Clinical and Experimental Pathology

Article Title: The protective effect of the Rho-kinase inhibitor hydroxyfasudil on propofol-induced hippocampal neuron apoptosis in neonatal rats

doi:

Figure Lengend Snippet: Hydroxyfasudil improves the tendency towards cognitive impairments induced by propofol. Evaluation of rat learning and memory in the Morris water maze. A. Mean escape latency to the platform in the Morris water maze. B. Time spent in platform quadrant among the different groups. C and D. Pull-down experiments for GTP-bound RhoA in primary culture hippocampal neurons with indicated treatment. Data were shown as the mean ± SD, **P < 0.01 vs. the control group; #P < 0.05, ##P < 0.01 vs. the “P” group.

Article Snippet: RhoA activation assay The RhoA activation assay was performed to detect the endogenous active form of RhoA (RhoA-GTP) using a RhoA activation assay kit (Abcam; ab211164) following the manufacturer’s manuals.

Techniques: