Fig 2: Expression of SGLT2 on mouse heart in infarcted area. (A) Representative images of immunofluorescence stain of SGLT2 on mouse heart 72 hours after MI. The infarcted zones are demarcated with dotted line. SGLT2 stained prominently on infarcted area regardless of EMPA treatment. (B) Quantitative analysis of SGLT2 fluorescence densitogram. Both vehicle and EMPA group showed significant increase in SGLT2 intensity. However, no difference between vehicle and EMPA group was noted (Scale bars: 500 μm).DAPI = 4′,6-diamidino-2-phenylindole; EMPA = empagliflozin; MI = myocardial infarction; SGLT2 = sodium/glucose cotransporter 2; Veh = vehicle.*p<0.05, †p<0.001 between the 2 groups connected by line.

Fig 3: The effects of uninephrectomy (UNx) and/or dapagliflozin administration on sodium‐glucose cotransporter (SGLT) protein, phosphoenolpyruvate carboxykinase (Pck1), and pyruvate kinase (Pklr) expression in the kidney tissue of SDT fatty rats. (a) Western blot analysis using antibodies against SGLT2 and β‐actin in the sham and UNx groups, (b) SGLT2/β‐actin protein abundance in the sham and UNx groups (mean ± standard deviation). (c) Pck1/β‐actin mRNA ratios in the four treatment groups (mean ± standard deviation). (d) Pklr/β‐actin mRNA ratios in the four treatment groups (mean ± standard deviation). Data were evaluated using one‐way ANOVA, with Bonferroni correction. ns: Not significant.

Fig 4: SGLT2 as a 1,5-AG transporter in the kidney.A Differentially expressed proteins of kidney tissues in UUO vs Sham are highlighted with volcano plot. The blue dots indicate the downregulated proteins, and the red dots indicate the upregulated proteins. B Western blot shows SGLT2 protein expression in kidneys from sham, UUO and UUO + BF (B. fragilis). C Quantitative analysis of panel B (n = 3). ***p = 0.0004 for SGLT2: Sham vs. UUO, **p = 0.0072 for SGLT2: Sham vs. UUO + BF, #p = 0.0394 for SGLT2: UUO vs. UUO + BF. D mRNA level of Slc5a2 was investigated in the GEO database using 153 subjects. Log-transformed SLC5A2 mRNA level was compared not only between glomeruli from healthy people (Normal) (n = 14) and lupus nephritic patients (LN) (n = 32), but also between tubulointerstitium from healthy people (n = 15) and lupus nephritic patients (n = 32) using Mann-Whitney U test; Log-transformed SLC5A2 mRNA level was compared not only between glomeruli from healthy people (n = 9) and hypertensive nephropathic patients (HN) (n = 9), but also between glomeruli from healthy people (n = 15) and IgA nephropathic patients (n = 27) using Mann-Whitney U test. **p = 0.0083 for Glomeruli HN; ***p < 0.0001 for Glomeruli LN; **p = 0.0096 for Glomeruli IgA; **p = 0.0091 for Tubulointerstitium LN. E mRNA level of SLC5A2 in the indicated groups (n = 6). ***p < 0.0001 for Slc5a2: Sham vs. UUO, *p = 0.0140 for SGLT2: Sham vs. UUO + BF, #p = 0.0124 for SGLT2: UUO vs. UUO + BF. F The root mean square deviation (RMSD) of protein backbone atoms and 1,5-AG during the molecular dynamics (MD) simulation. G, H Molecular docking and the binding mode of 1,5-AG to SGLT2 through the MD simulation. I Empagliflozin significantly decreased the serum concentration of 1,5-AG in control mice (n = 10). ***p = 0.0003. Box plots show center line as median, whiskers show maxima and minima, and box limits show upper and lower quartiles. J Cellular uptake experiments of 1,5-AG-13C6 were performed in stably SGLT2 transfected HEK293 vs wide-type (WT) cells (n = 6). ***p = 0.0022. Data are presented as mean ± SD. Comparison in D was performed with a two-tailed Mann-Whitney U test. Comparison in I and J were performed with a two-tailed Student’s t test. Comparisons in C, E were compared using One-Way ANOVA followed by Sidak’s multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001 (compared with sham group or normal subjects), #P < 0.05, ##P < 0.01, ###P < 0.001 (compared with UUO group). Individual data points are independent biological replicates unless otherwise stated.

Fig 5: Canagliflozin inhibits cancer cell proliferation. (A–C) Cancer cell proliferation with SGLT2 inhibitors (canagliflozin, dapagliflozin) or metformin in (A) SKBR3 cells (n = 4), (B) BT-474 cells (n = 4), and (C) NT2197 cells (n = 3). (D) Cell proliferation of SKBR3 cells treated with SGLT2 inhibitors (canagliflozin, dapagliflozin) with and without metformin, or DMSO (control), and grown in media containing glucose (11 mM) or galactose (11 mM) for 48 h (n = 4). (E) Cell viability (%) of the cells grown in (D). (F) Knockdown of SGLT2 in SKBR3 cells (n = 3). Western blot of SGLT2 and β-Actin expression in siCTRL and siSLC5A2 SKBR3 cells (n = 3). All data are presented as means + SEM, *P< 0.05. (A-C) One-way ANOVA, Dunnett's posthoc test with untreated cells as control; (D-E) Two-way ANOVA, Dunnett's posthoc test with untreated cells grown in glucose as control; (F) Two-way ANOVA, Tukey's posthoc test.