Fig 2: Expression of SGLT2 in Hep3B and Huh7.A) Immunoblotting using 4 different antibodies for SGLT2: Antibody-a, Proteintech Group, Inc. (24654-1-AP); Antibody-b, Abcam plc. (ab137207); Antibody-c, Abcam plc. (ab85626), Antibody-d, Cell Signaling Technology, Inc. (#14210). B) Immunohistochemistry for SGLT2. Green indicates SGLT2. The nucleus was stained by DAPI. Scale bar = 50 μm. C) Double immunostaining for mitochondrial PDK-1 and SGLT2. Green indicates mitochondrial PDK-1. Red indicates SGLT2. Yellow indicates colocalization of mitochondrial PDK-1 and SGLT2. The nucleus was stained by DAPI. Scale bar = 50 μm. D) Immunoblotting for SGLT2 in mitochondrial and cytoplasmic fractions. Abbreviations: SGLT2, sodium-glucose cotransporter 2; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; DAPI, 4',6-diamidino-2-phenylindole; PDK-1, pyruvate dehydrogenase kinase 1.

Fig 3: Schematic model depicting SGLT2 inhibitor-mediated acquisition of ischemic tolerance in brain pericytes.Luseogliflozin, an SGLT2 inhibitor, inhibits Na+-dependent glucose uptake and activates AMPKα in response to depletion of intracellular glucose and ATP. Activated AMPKα enhances mitochondrial biogenesis via PGC-1α, NRF1, and NRF2 signaling, leading to ischemic tolerance acquisition. SGLT2 sodium-glucose cotransporter 2, TFAM mitochondrial transcription factor A.

Fig 4: Survival from MACE Cox regression analysis (adjusted for age, sex, BMI, blood pressure, heart rate, cholesterol, HDL cholesterol, LDL cholesterol, triglyceride levels, heart disease, hypertension, dyslipidemia, smoking, b-blockers, ACE inhibitors, calcium inhibitors, thiazide diuretics, and aspirin) according to diabetic status (A), SGLT2i therapy (B), and SGLT2 carotid atherosclerotic plaque content (C).

Fig 5: SGLT2 expression in cultured pericytes is upregulated by glucose and/or oxygen deprivation.a Cultured brain pericytes were treated with GD for 16 h, OD for 16 h, OGD for 16 h, or OGD followed by 24-h reperfusion (rep.) with normal glucose and oxygen level. The mRNA expression level of SLC5A2 was quantified using qPCR and normalized to that of RNA18S (n = 5). b Representative images of immunofluorescence for SGLT2 (green) and DAPI (blue) in pericytes treated with or without OGD for 16 h (scale bar = 20 μm). Data are presented as dot-plots of individual experiments and mean values ± SD. n.s. (not significant) P > 0.05, ****P < 0.0001 by One-way ANOVA followed by Bonferroni’s post hoc test. SGLT2 sodium-glucose cotransporter 2, Luseo luseogliflozin, GD glucose deprivation, OD oxygen deprivation, OGD oxygen-glucose deprivation.