Journal: Stem Cell Research & Therapy

Article Title: HMGB1-mediated enhancement of glycolysis activates hepatic stellate cells by inhibiting ferroptosis in alcoholic hepatic fibrosis

doi: 10.1186/s13287-026-04970-1

Figure Lengend Snippet: Elevated ethanol concentration influenced the metabolism of HSCs. A – D The effects of ethanol concentration on glucose consumption and the production of pyruvate, ATP and lactate in ethanol-stimulated LX-2 cells were assessed by kit assays. E WB assay was used to detect the effect of ethanol concentration on the expression levels of HK2, PKM2, LDHA and PDH in ethanol-stimulated LX-2 cells. F The effects of ethanol concentration on the activities of HK, PK, LDH and PHD in LX-2 cells stimulated by ethanol were measured by kit detections. G – H Seahorse XF assay was utilized to observe the effect of ethanol concentration on OCR and ECAR of ethanol-stimulated LX-2 cells. * p ˂ 0.05, ** p ˂ 0.01, *** p ˂ 0.001 vs. 0 µg/mL Ethanol

Article Snippet: After deparaffinization and rehydration, tissue slides were treated with 100 μL endogenous peroxidase blockers for 10 min. Then, the slides were incubated with polyclonal primary antibody against HMGB1 (A00066-1, 1:200; Boster, Pleasanton, CA, USA), glutathione peroxidase 4 (GPX4) (BM5231, 1:200; Boster), ferritin heavy chain 1 (FTH1) (ab183781, 1:500; Abcam, Cambridge, MA, USA), nuclear factor erythroid 2-related factor 2 (NRF2) (A0674, 1:200; Abclonal, Beijing, China), pyruvate kinase M2 (PKM2) (PB9379, 1:200; Boster), lactate dehydrogenase A (LDHA) (PB10075, 1:200; Boster), Collagen IV (ab6586, 1:500; Abcam), and alpha-smooth muscle actin (α-SMA) (BM4172, 1:200; Boster) overnight at 4 °C.

Techniques: Concentration Assay, Expressing, XF Assay

Journal: Stem Cell Research & Therapy

Article Title: HMGB1-mediated enhancement of glycolysis activates hepatic stellate cells by inhibiting ferroptosis in alcoholic hepatic fibrosis

doi: 10.1186/s13287-026-04970-1

Figure Lengend Snippet: HMGB1 regulated HSC activation and ECM deposition by modulating its metabolism. A – D The effects of si-HMGB1 on glucose consumption and the production of pyruvate, ATP and lactate in ethanol-stimulated LX-2 cells were evaluated by kit assays. E WB assay was employed to detect the effect of si-HMGB1 on the expression levels of HK2, PKM2, LDHA and PDH in ethanol-stimulated LX-2 cells. F The effects of si-HMGB1 on the activities of HK, PK, LDH and PHD in ethanol-stimulated LX-2 cells were assessed by kit assays. G – I Seahorse XF assay was used to observe the effect of si-HMGB1 on OCR and ECAR of ethanol-stimulated LX-2 cells. J – K The effects of OLIG and 2-DG on the regulation of OCR and ECAR by si-HMGB1 or HMGB1 overexpression in alcohol-stimulated LX-2 cells were measured by Seahorse XF assays. L – M CCK-8 and EdU staining assays were utilized to evaluate the effect of OLIG and 2-DG on si-HMGB1 or HMGB1 overexpression regulating the viability and proliferation of alcohol-stimulated LX-2 cells. N The effects of OLIG and 2-DG on the ECM of alcohol-stimulated LX-2 cells regulated by si-HMGB1 or HMGB1 overexpression were examined by WB. * p ˂ 0.05, ** p ˂ 0.01, *** p ˂ 0.001 vs. Ethanol/ Ethanol + si-NC/ Ethanol + si-HMGB1/ Ethanol+Vector/ Ethanol+HMGB1

Article Snippet: After deparaffinization and rehydration, tissue slides were treated with 100 μL endogenous peroxidase blockers for 10 min. Then, the slides were incubated with polyclonal primary antibody against HMGB1 (A00066-1, 1:200; Boster, Pleasanton, CA, USA), glutathione peroxidase 4 (GPX4) (BM5231, 1:200; Boster), ferritin heavy chain 1 (FTH1) (ab183781, 1:500; Abcam, Cambridge, MA, USA), nuclear factor erythroid 2-related factor 2 (NRF2) (A0674, 1:200; Abclonal, Beijing, China), pyruvate kinase M2 (PKM2) (PB9379, 1:200; Boster), lactate dehydrogenase A (LDHA) (PB10075, 1:200; Boster), Collagen IV (ab6586, 1:500; Abcam), and alpha-smooth muscle actin (α-SMA) (BM4172, 1:200; Boster) overnight at 4 °C.

Techniques: Activation Assay, Expressing, XF Assay, Over Expression, CCK-8 Assay, Staining, Plasmid Preparation