Fig 2: Hepatic PTP1B disruption mitigates ethanol-induced injury and inflammation. Ctrl (Ptpn1fl/fl) and KO (Ptpn1fl/fl, Alb-Cre) female mice were used in the chronic plus binge model. Pair: control diet + maltose gavage, EtOH: ethanol diet + ethanol gavage. A) Plasma ALT presented in a bar chart as means + SEM (n = 5 pair-fed Ctrl mice and n = 6 for each of the remaining groups). **p < 0.01 Pair vs. EtOH and ††p < 0.01 Ctrl vs. KO by one-way ANOVA with post-hoc Tukey's test. B) Hepatic mRNA of Il1b, Ccl2, Tnfa, Adgre1, and Ly6g were determined by qPCR normalized to Tbp then expressed as means + SEM (n = 3 per group). *p < 0.05 Pair vs. EtOH and †p < 0.05 Ctrl vs. KO by a two-tailed t-test. C) H&E-stained liver sections of Ctrl and KO mice with ethanol-induced inflammatory cell infiltration indicated by white circles. Scale bar: 100 µm. D) Confocal images of liver sections immunostained with the macrophage marker F4/80 (green) and DAPI counterstain (blue). Boxed areas are enlarged, and arrowheads indicate macrophages. Scale bar: 50 µm. E) Immunoblots of pNF-?B and NF-?B in hepatic lysates and each lane represents an independent animal. The phosphorylation level was normalized to protein expression and represented as means + SEM (right panel; n = 5 pair-fed Ctrl mice and n = 6 for each of the remaining groups). *p < 0.05 Pair vs. EtOH and ††p < 0.01 Ctrl vs. KO by a two-tailed t-test. A.U.: arbitrary unit.

Fig 3: Hepatic PTP1B disruption attenuates ethanol-induced steatosis. Ctrl (Ptpn1fl/fl) and KO (Ptpn1fl/fl, Alb-Cre) female mice were used in the chronic plus binge model. Pair: control diet + maltose gavage, EtOH: ethanol diet + ethanol gavage. A) Hepatic triglycerides concentrations presented as means + SEM (n = 5 pair-fed Ctrl mice and n = 6 for each of the remaining groups). *p < 0.05, **p < 0.01 Pair vs. EtOH and †p < 0.05 Ctrl vs. KO by one-way ANOVA with post-hoc Tukey's test. B) Hepatic mRNA of Pparg, Srebf1, Fasn, Cd36, and Scd were determined by qPCR normalized to Tbp then expressed as means + SEM (n = 3 per group). *p < 0.05, **p < 0.01 Pair vs. EtOH and †p < 0.05, ††p < 0.01 Ctrl vs. KO by a two-tailed t-test. A.U.: arbitrary unit. C) H&E-stained liver sections of Ctrl and KO mice demonstrating lipid accumulation and boxed areas are enlarged. Scale bar: 100 µm.

Fig 4: Detection of oxidized PTP1B by dimedone-PLA. Serum-starved HepG2 cells expressing control shRNA or shRNA targeting human PTPN1 (shPTPN1), with or without stable expression of either mouse WT PTP1B or PTP1BC215S, were stimulated with insulin (25 nM) or H2O2 (1 mM) for 5 min or left unstimulated. Cells were then subjected to dimedone-PLA with anti-dimedone-Cys and anti-mouse PTP1B antibodies (gray). Representative images are shown for each condition from one of two independent biological replicates. The graph shows the average number of PLA signals per cell (n = 6 images for each condition, 80–120 cells in an image), relative to control cells without stimulation (set as 1). ***P < 0.0001, ns not significant, ANOVA with Bonferroni/Dunn’s post-hoc test. Error bars represent SD. Scale bar: 50 µm

Fig 5: PTP1B pharmacological inhibition attenuates ethanol-induced hepatic injury, inflammation, steatosis, and oxidative stress. A) Schematic depicting the administration of PTP1B inhibitor (PTP1Bi; DPM-1001) in the chronic plus binge model. Wild-type female mice were provided 5% EtOH liquid diet without (white arrow) and with PTP1B inhibitor (black arrow) for ten days. Mice were sacrificed 9 h post oral gavage of 31.5% EtOH (0.02 ml/g body weight) on day 11. (B) Plasma ALT level (left panel; n = 6 EtOH and n = 7 EtOH + PTP1Bi) and hepatic mRNA of Il1b, Ccl2 and Tnfa (right panel; n = 4 per group), and (C) hepatic triglycerides (TGs; n = 7 EtOH and n = 8 EtOH + PTP1Bi) and mRNA of Fasn, Pparg and Srebf1 (right panel; n = 4 each group) are presented as means + SEM. *p < 0.05, **p < 0.01 EtOH vs. EtOH + PTP1Bi by a two-tailed t-test. D) Immunoblots of pNF-?B, NF-?B, pAKT, AKT, pERK, ERK, NOX2, NOX4, SOD1, GPx1/2, and GAPDH in mouse liver lysates. Each lane represents an independent animal.