Obtained pO₂ values in tissue near the focus and surround.

<p>(a) Grayscale angiogram of cortical surface and locations for pO₂ measurement (red: focus; blue: surround). The artery was shown by the red arrows. Scale bar size: 0.2mm (b) Epileptic activity induced a transient dip in tissue pO₂ followed by an increase in pO₂ in the focus. A sustained increase in pO₂ was seen in the surround. The dashed vertical lines show the ictal onset (left) and offset (right). (c) Distribution of percent of initial dip at multiple locations (color dotted) during epileptic activity. The 4-AP injection site is shown by green circle. The artery was shown by the red arrows. Scale bar size: 0.2mm.</p

(a) Grayscale angiogram of cortical pial tissue with points of interest (red dots). Scale bar size: 0.2mm (b) Corresponding temporal profiles of pO₂ measured while altering FiO₂. The gray segments denote the 10 minutes period during which FiO₂ was increased up to 40%.

<p>(a) Grayscale angiogram of cortical pial tissue with points of interest (red dots). Scale bar size: 0.2mm (b) Corresponding temporal profiles of pO₂ measured while altering FiO₂. The gray segments denote the 10 minutes period during which FiO₂ was increased up to 40%.</p

(a) Schematic of the confocal lifetime imaging system. Excitation light is provided by a laser diode (λ = 637nm, 170 mW maximum power, which is collimated by a convex lens (L1) and travels through the objective for illumination. It is focused onto the cranial window by a 10×magnification objective (Obj), which is directed to the specific points using galvanometric scanners (xy). Emitted phosphorescence light is separated from excitation light using a beam splitter (BS2) and filter (F) and detected with an avalanche photodiode (APD). The system is controlled by a computer through a data acquisition card (DAQ). (b) In vivo measurements of pO₂vs mean counts per millisecond as controlled by the diode laser power. Higher laser powers correlate with higher consumption of O₂ leading to a significant decrease of pO₂ estimates over time (seen in the first point when average counts exceed 10000). When limiting to 3000 average counts, no significant decrease in pO

<p>(a) Schematic of the confocal lifetime imaging system. Excitation light is provided by a laser diode (λ = 637nm, 170 mW maximum power, which is collimated by a convex lens (L1) and travels through the objective for illumination. It is focused onto the cranial window by a 10×magnification objective (Obj), which is directed to the specific points using galvanometric scanners (xy). Emitted phosphorescence light is separated from excitation light using a beam splitter (BS2) and filter (F) and detected with an avalanche photodiode (APD). The system is controlled by a computer through a data acquisition card (DAQ). (b) In vivo measurements of pO₂vs mean counts per millisecond as controlled by the diode laser power. Higher laser powers correlate with higher consumption of O₂ leading to a significant decrease of pO₂ estimates over time (seen in the first point when average counts exceed 10000). When limiting to 3000 average counts, no significant decrease in pO₂ could be measured over time. (c) Example of phosphorescence decay profiles under conditions where photo-consumption is negligible. Higher O₂ concentration causes more quenching of phosphorescence signal, and consequently a faster decay (red profile).</p

Temperature dependencies of oxygen quenching constants (Kq) and lifetimes (τ0) for G4 (a and b).

<p>The measurements were performed using 50 μM solutions of the probes, pH 7.23.</p

Measured pO₂ values during normoxia (color dots), overlaid with a grayscale angiogram of cortical pial tissue from an exposed window(with an artery shown by the red arrows).

<p>The size of scale bar is 0.2mm.</p

Electrophysiology of 4-AP induced epileptic activity.

<p>Top: example of ictal discharges after the 4-AP injection. Middle: zoom on an ictal discharge. Bottom: expanded view of showing the onset of the discharge, the intermediate phase and the offset.</p

a) Correlation between initial dip (% change) and duration of epileptic activity. The line of linear fit was <i>y</i> = 0.74<i>x</i> − 4.35, R² = 0.81 (b) Statistical distribution of the slopes for all mice. M1 was the name of mouse and number in the bracket was the number of seizure that was calculated. The outliers were plotted with red plus sign. The average of goodness of fit (R2) was listed for each mouse.

<p>a) Correlation between initial dip (% change) and duration of epileptic activity. The line of linear fit was <i>y</i> = 0.74<i>x</i> − 4.35, R² = 0.81 (b) Statistical distribution of the slopes for all mice. M1 was the name of mouse and number in the bracket was the number of seizure that was calculated. The outliers were plotted with red plus sign. The average of goodness of fit (R2) was listed for each mouse.</p

mTORC2 loss exacerbates the pro-inflammatory profile of BMDM in response to several TLR agonists.

<p>(A to D) Impact of several TLR agonists on gene expression in macrophages isolated from LysM-<i>Rictor</i>^WT or LysM-<i>Rictor</i>^KO mice. BMDM were isolated from LysM-<i>Rictor</i>^WT or LysM-<i>Rictor</i>^KO mice and were differentiated for 6 days <i>in vitro</i>. Cells were then plated and incubated 24 hours. Cells were then treated with (A) PAM3 (1ug/ml), (B) MALP2 (0.1ug/ml), (C) R848 (0.1uM), or CpG (0.5uM) for 8 hours. mRNA expression of markers of the classically activated (M1) and alternatively activated (M2) macrophages were measured by qRT-PCR and normalized to 36B4 mRNA levels. Data are expressed as the mean ± SEM for n = 4 per condition. *p<0.05 versus control. This experiment was repeated twice and similar results were observed.</p

LysM-<i>Rictor</i>^KO mice exhibit a higher susceptibility to LPS-induced septic shock.

<p>LysM-<i>Rictor</i>^WT and LysM-<i>Rictor</i>^KOmice were injected intraperitoneally with LPS (2.5 mg/kg of body weight) and body temperature was measured 4 hours post-injection. * denotes a significant difference between LysM-<i>Rictor</i>^WT or LysM-<i>Rictor</i>^KO mice (p<0.05). Data are expressed as the mean ± SEM for n = 6–10 per condition. (B) Survival rate of LysM-<i>Rictor</i>^WT or LysM-<i>Rictor</i>^KO mice following the injection of LPS. Mice having a core body temperature dropping down to 33.0+/−0.2°C or below following LPS injection were killed. (C) The sensitivity of LysM-<i>Rictor</i>^KO mice to LPS-induced hypothermia is linked to an elevation in circulating levels of TNF-α. Plasma was collected from the mice used in the experimentation described in A before LPS injection and 1,2, and 4 hours post injection. TNF-α levels were measured by ELISA. Data are expressed as the mean ± SEM. The graph presented in B is based on the plasma measurements of TNF-α of all LysM-<i>Rictor</i>^WT mice (n = 6) vs. the LysM-<i>Rictor</i>^KO mice that died over the experiment (n = 7). *p<0.05 versus control.</p

Myeloid-specific deletion of <i>Rictor</i> does not affect metabolic homeostasis in mice fed a low or a high-fat diet.

<p>(A–D) Body and tissue weight of LysM-<i>Rictor</i>^WT or LysM-<i>Rictor</i>^KO mice fed a (A and C) low or (B a D) high fat diet for 21 and 23 weeks respectively. (E) Blood parameters of LysM-<i>Rictor</i>^WT or LysM-<i>Rictor</i>^KO mice fed either a low or a high fat diet. Mice were fasted for 6 hours before blood collection. ^Ŧ denotes a significant difference between low fat fed and high fat fed mice (p<0.05). The p value for the genotype effect is indicated on the right side of the table. (F–G) GTT and ITT of LysM-<i>Rictor</i>^WT or LysM-<i>Rictor</i>^KO mice fed with (F) low or (G) high fat diet. For the panels A to G, n = 6–13 mice per group. (H–I) Gene expression in (H) WAT and (I) liver of LysM-<i>Rictor</i>^WT or LysM-<i>Rictor</i>^KO mice fed a low or a high fat diet. mRNA expression was measured by qRT-PCR and normalized to 36B4 mRNA levels. Data are expressed as the mean ± SEM for n = 6–10 per condition. ^Ŧ denotes a significant difference between low fat fed and high fat fed mice (p<0.05). * denotes a significant difference between LysM-<i>Rictor</i>^WT or LysM-<i>Rictor</i>^KO mice (p<0.05).</p