Journal: Cells

Article Title: Canagliflozin, an Inhibitor of the Na + -Coupled D-Glucose Cotransporter, SGLT2, Inhibits Astrocyte Swelling and Brain Swelling in Cerebral Ischemia.

doi: 10.3390/cells12182221

Figure Lengend Snippet: Figure 2. SGLT2 is upregulated in astrocyte cell bodies and astrocyte endfeet following cerebral ischemia. (a–e) Double immunolabeling for SGLT2 (green) and NeuN (a,b) or GFAP (c–e) (red) in contralateral (Contra) and ipsilateral (Ipsi) brain sections from mice post-MCAo/R (2/6 h); merged images are shown in a, b, d, e; individual labelings are shown in c, d—inset, e—inset; the images shown are representative of findings in 3 mice; scale bars, 25 µm except insert, 10 µm.

Article Snippet: The immunoblot of the kidney lysate showed a pair of bands, ~60 and ~65 kDa, with a third band, ~26 kDa, which were absent or minimal in lung lysate and were blocked by pre-absorption of the anti-SGLT2 antibody with the recombinant SGLT2 antigen peptide (#NBP1-92384PEP; Novus Biologicals) (Figure S3a).

Techniques: Immunolabeling

Journal: Cells

Article Title: Canagliflozin, an Inhibitor of the Na + -Coupled D-Glucose Cotransporter, SGLT2, Inhibits Astrocyte Swelling and Brain Swelling in Cerebral Ischemia.

doi: 10.3390/cells12182221

Figure Lengend Snippet: Figure 3. Slc5a2 mRNA and SGLT2 protein are upregulated in astrocytes following cerebral ischemia. (a–c) Images (a) and quantification (b,c) of RNAscope for Slc5a2 (red) and Aqp4 (green) mRNA in contralateral (Contra) and ipsilateral (Ipsi) brain sections from mice post-MCAo/R (2/6 h); scale bar, 25 µm; data from 3 mice per group; **, p < 0.01. (d) qPCR for Slc5a2 mRNA in astrocytes and neurons isolated from the contralateral (Contra) and ipsilateral (Ipsi) MCA territory of mice following MCAo/R (2/6 h) and uninjured controls (naïve), expressed as fold-change; 5 mice per group; *, p < 0.05. (e) Quantitative immunohistochemistry for SGLT2 expressed within regions of interest determined by NeuN; data from 7 mice per group; ns, not significant. (f,g) Immunoblot (f) and quantification of immunoblot (g) for SGLT2 in astrocytes isolated from the contralateral (Contra) and ipsilateral (Ipsi) MCA territory of mice following MCAo/R (2/6 h) and uninjured controls (naïve), expressed as fold-change; PC, positive control (kidney lysate); β-actin used as a loading control; 3 mice per group.

Article Snippet: The immunoblot of the kidney lysate showed a pair of bands, ~60 and ~65 kDa, with a third band, ~26 kDa, which were absent or minimal in lung lysate and were blocked by pre-absorption of the anti-SGLT2 antibody with the recombinant SGLT2 antigen peptide (#NBP1-92384PEP; Novus Biologicals) (Figure S3a).

Techniques: RNAscope, Isolation, Immunohistochemistry, Western Blot, Positive Control, Control

Journal: Cells

Article Title: Canagliflozin, an Inhibitor of the Na + -Coupled D-Glucose Cotransporter, SGLT2, Inhibits Astrocyte Swelling and Brain Swelling in Cerebral Ischemia.

doi: 10.3390/cells12182221

Figure Lengend Snippet: Figure 4. D-glucose-induced Na+ influx in post-ischemic astrocytes is inhibited by canagliflozin and by the astrocyte-specific deletion of Slc5a2/SGLT2. (a) Live cell image of a tdTomato-expressing astrocyte in an ex vivo brain slice from an MCAo/R (2/6 h) mouse, following incubation with ING-2; scale bar, 25 µm. (b–e) Time course of Na+ influx with step change in D-glucose from 2 to 10 mM in ipsilateral (Ipsi) vs. contralateral (Contra) astrocytes (b); in ipsilateral astrocytes without and with pre-incubation with canagliflozin (5 µM) (c); in ipsilateral astrocytes without and with the addition of canagliflozin (5 µM) at the time indicated by the bar (d); in ipsilateral astrocytes from Ast-Slc5a2WT

Article Snippet: The immunoblot of the kidney lysate showed a pair of bands, ~60 and ~65 kDa, with a third band, ~26 kDa, which were absent or minimal in lung lysate and were blocked by pre-absorption of the anti-SGLT2 antibody with the recombinant SGLT2 antigen peptide (#NBP1-92384PEP; Novus Biologicals) (Figure S3a).

Techniques: Expressing, Ex Vivo, Slice Preparation, Incubation