Journal: Molecular Metabolism

Article Title: Hepatic stellate cell-specific Kcnma1 deletion mitigates metabolic dysfunction-associated steatotic liver disease progression via upregulating Amphiregulin secretion

doi: 10.1016/j.molmet.2025.102164

Figure Lengend Snippet: Kcnma1 knockout protects against HFD-induced MASLD. ( A and B ) Weight growth curve and body weight gain of Kcnma1 WT mice and Kcnma1 KO mice after 12 weeks HFD administration ( n = 10–11 biologically independent mice per condition). ( C ) Representative image of liver appearance alteration in HFD-treated WT and Kcnma1 KO mice (Scale bar, 1 cm). ( D ) Liver weight of Kcnma1 WT mice and Kcnma1 KO mice after 12 weeks HFD administration ( n = 5 biologically independent mice per condition). ( E-H ) Representative images of epididymal white adipose tissue (eWAT) and inguinal white adipose tissue (iWAT) from Kcnma1 WT and knockout KO mice after HFD treatment, along with statistical analysis of their tissue weights ( n = 8 biologically independent mice per condition) (Scale bar, 1 cm). ( I ) Liver Triglycerides (TG) contents of Kcnma1 WT mice and Kcnma1 KO mice after HFD for 12 weeks. ( J-O ) Triglycerides (TG), total cholesterol (TC), low-density lipoprotein (LDL) and high-density lipoprotein (HDL), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in the serum were quantified in Kcnma1 WT and Kcnma1 KO mice ( n = 5 biologically independent mice per condition). ( P and Q ) Representative H&E staining and Oil Red O staining images of the liver sections from the indicated groups ( n = 5/group) (Scale bar, 100 μm). ( R and S ) GTT and ITT assays of HFD-fed Kcnma1 WT mice and Kcnma1 KO mice ( n = 6 biologically independent mice per condition). ns, not significant; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001 (unpaired two-tailed Student’s t test; mean ± SEM). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The following day, the membrane was incubated with a horseradish peroxidase (HRP)-conjugated secondary antibody for 1 h. The antibodies for immunoblots including KCNMA1 (75-022, L6/60, NeuroMab, Mouse/IgG, 1:1000), AREG (A1860, ABclonal, Rabbit/IgG, 1:1000), Erk (4695, CST, Rabbit/IgG, 1:1000), p-Erk (4370, CST, Rabbit/IgG, 1:1000), EGFR (4267, CST, Rabbit/IgG, 1:1000), GAPDH (2118, CST, Rabbit/IgG, 1:1000) and Tubulin (T0023, Affnity Biosciences, Mouse/IgG, 1:1000).

Techniques: Knock-Out, Staining, Two Tailed Test

Journal: Molecular Metabolism

Article Title: Hepatic stellate cell-specific Kcnma1 deletion mitigates metabolic dysfunction-associated steatotic liver disease progression via upregulating Amphiregulin secretion

doi: 10.1016/j.molmet.2025.102164

Figure Lengend Snippet: Kcnma1 in hepatocytes is irrelevant to liver metabolic disorders induced by HFD. ( A and B ) Weight growth curve and body weight gain of Kcnma1 -Flox mice and Kcnma1 -HKO mice after 12 weeks HFD treatment ( n = 11 biologically independent mice per condition). ( C ) Representative image of liver appearance alteration in HFD-treated Kcnma1 -Flox mice and Kcnma1 -HKO mice (Scale bar, 1 cm). ( D ) Liver weight of Kcnma1 -Flox mice and Kcnma1 -HKO mice after 12 weeks HFD treatment ( n = 6 biologically independent mice per condition). (E) Liver triglycerides (TG) contents of Kcnma1 -Flox mice and Kcnma1 -HKO mice after HFD for 12 weeks ( n = 6 biologically independent mice per condition). ( F ) Representative H&E staining and Oil Red O staining images of the liver sections from the indicated groups ( n = 5–6 biologically independent mice per condition). ( G ) TG, total cholesterol (TC), low-density lipoprotein (LDL) and high-density lipoprotein (HDL), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in the serum were determined in Kcnma1 -Flox mice and Kcnma1 -HKO mice ( n = 6 biologically independent mice per condition). ns, not significant (unpaired two-tailed Student’s t test; mean ± SEM). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The following day, the membrane was incubated with a horseradish peroxidase (HRP)-conjugated secondary antibody for 1 h. The antibodies for immunoblots including KCNMA1 (75-022, L6/60, NeuroMab, Mouse/IgG, 1:1000), AREG (A1860, ABclonal, Rabbit/IgG, 1:1000), Erk (4695, CST, Rabbit/IgG, 1:1000), p-Erk (4370, CST, Rabbit/IgG, 1:1000), EGFR (4267, CST, Rabbit/IgG, 1:1000), GAPDH (2118, CST, Rabbit/IgG, 1:1000) and Tubulin (T0023, Affnity Biosciences, Mouse/IgG, 1:1000).

Techniques: Staining, Two Tailed Test

Journal: Molecular Metabolism

Article Title: Hepatic stellate cell-specific Kcnma1 deletion mitigates metabolic dysfunction-associated steatotic liver disease progression via upregulating Amphiregulin secretion

doi: 10.1016/j.molmet.2025.102164

Figure Lengend Snippet: Hepatic stellate cells specific deletion of Kcnma1 mitigates hepatic steatosis in HFD-fed mice. ( A ) UMAP plots of the subtypes cells in mice livers and Kcnma1 -positive cell types. Each cluster is color-coded according to cell type. Cluster annotations are indicated in figure. ( B ) Schematic of Kcnma1 specific knockout in hepatic stellate cells. ( C ) PCR and western blotting analysis of Kcnma1 expression in primary hepatic stellate cells. ( D and E ) Weight growth curve and body weight gain of Kcnma1 -Flox mice and Kcnma1 -HSCKO mice after 12 weeks HFD administration ( n = 7 biologically independent mice per condition). ( F ) Representative image of liver appearance alteration in HFD-treated Kcnma1 -Flox mice and Kcnma1 -HSCKO mice. (scale bar, 1 cm). ( G ) Liver weight and of Kcnma1 -Flox mice and Kcnma1 -HSCKO mice after 12 weeks HFD administration ( n = 5–7 biologically independent mice per condition). ( H ) Determination of liver TG contents of Kcnma1 -Flox mice and Kcnma1 -HSCKO mice after HFD for 12 weeks ( n = 7–8 biologically independent mice per condition). ( I and J ) Representative image of H&E staining and Oil Red O staining images of the liver sections from the indicated groups ( n = 9 biologically independent mice per condition) (Scale bar, 100 μm). ( K–P ) Triglycerides (TG), total cholesterol (TC), low-density lipoprotein (LDL) and high-density lipoprotein (HDL), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the serum were determined in Kcnma1 -Flox mice and Kcnma1 -HSCKO mice ( n = 5 biologically independent mice per condition). ( Q and R ) GTT and ITT assays of HFD-fed Kcnma1 -Flox mice and Kcnma1 -HSCKO mice ( n = 7 biologically independent mice per condition). ns, not significant; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001 (unpaired two-tailed Student’s t test; mean ± SEM). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The following day, the membrane was incubated with a horseradish peroxidase (HRP)-conjugated secondary antibody for 1 h. The antibodies for immunoblots including KCNMA1 (75-022, L6/60, NeuroMab, Mouse/IgG, 1:1000), AREG (A1860, ABclonal, Rabbit/IgG, 1:1000), Erk (4695, CST, Rabbit/IgG, 1:1000), p-Erk (4370, CST, Rabbit/IgG, 1:1000), EGFR (4267, CST, Rabbit/IgG, 1:1000), GAPDH (2118, CST, Rabbit/IgG, 1:1000) and Tubulin (T0023, Affnity Biosciences, Mouse/IgG, 1:1000).

Techniques: Knock-Out, Western Blot, Expressing, Staining, Two Tailed Test

Journal: Molecular Metabolism

Article Title: Hepatic stellate cell-specific Kcnma1 deletion mitigates metabolic dysfunction-associated steatotic liver disease progression via upregulating Amphiregulin secretion

doi: 10.1016/j.molmet.2025.102164

Figure Lengend Snippet: Kcnma1 disruption of HSCs reduces intracellular TG contents of hepatocytes cultured in 2D and 3D spheroids. ( A ) Assay of TG content in primary mouse hepatocytes treated with CM from OA/PA-treated primary mouse WT and Kcnma1 -KO HSCs, as well as control CM. ( B ) Assay of TG content in HepG2 treated with CM from OA/PA-treated WT and Kcnma1 -KO LX-2, as well as control CM. ( C ) Representative image of BODIPY493/503 lipid droplet staining of primary mouse hepatocytes induced by CM from OA/PA-treated primary mouse WT and Kcnma1 -KO HSCs. (Scale bar, 25 μm). ( D ) Representative image of BODIPY493/503 lipid droplet staining of HepG2 induced by CM from OA/PA-treated WT and Kcnma1 -KO LX-2 (Scale bar, 25 μm). ( E ) Representative immunofluorescence images of 3D spheroids (Hepatocyte/LX-2, 4:1) stained for Albumin (green), Lrat (red) and nuclei (DAPI, blue) (Scale bar, 100 μm). ( F ) Intracellular neutral lipid droplet in HepG2 of 3D spheroids visualized by BODIPY493/503 staining. (Scale bars, 100 μm). ( G ) Measurement of TG content in HepG2 cells treated with CM from Kcnma1 -overexpression (OE) LX-2 and control. ( H ) Representative images of BODIPY493/503 lipid droplet staining in hepatocytes treated with CM from Kcnma1 -OE LX-2 and control (Scale bar, 25 μm). ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001 (unpaired two-tailed Student’s t test; mean ± SEM). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The following day, the membrane was incubated with a horseradish peroxidase (HRP)-conjugated secondary antibody for 1 h. The antibodies for immunoblots including KCNMA1 (75-022, L6/60, NeuroMab, Mouse/IgG, 1:1000), AREG (A1860, ABclonal, Rabbit/IgG, 1:1000), Erk (4695, CST, Rabbit/IgG, 1:1000), p-Erk (4370, CST, Rabbit/IgG, 1:1000), EGFR (4267, CST, Rabbit/IgG, 1:1000), GAPDH (2118, CST, Rabbit/IgG, 1:1000) and Tubulin (T0023, Affnity Biosciences, Mouse/IgG, 1:1000).

Techniques: Disruption, Cell Culture, Control, Staining, Immunofluorescence, Over Expression, Two Tailed Test

Journal: Molecular Metabolism

Article Title: Hepatic stellate cell-specific Kcnma1 deletion mitigates metabolic dysfunction-associated steatotic liver disease progression via upregulating Amphiregulin secretion

doi: 10.1016/j.molmet.2025.102164

Figure Lengend Snippet: Paracrine AREG from Kcnma1 KO HSCs subsides lipid accumulation during MASLD . ( A ) Schematic procedure of cytokine array used in the CM transfer culture model of primary mouse cells. ( B ) Dot plot of expression levels of cytokine in array. Dot plot displaying average scaled expression levels of indicated proteins. Dot size reflected the relative expression level of respective proteins. ( C ) Determination of AREG levels in CM of primary mouse HSCs treated with FFAs by ELISA. ( D ) Determination of AREG levels in CM of LX-2 treated with FFAs by ELISA. ( E ) Measurement of TG levels in HepG2 cells treated with CM from Kcnma1 -knockdown LX-2 cells, Areg -knockdown LX-2 cells, and their respective controls. ( F ) Analysis of TG content in HepG2 treated with FFAs and 500 ng/ml AREG for 24 h, respectively. ( G ) Oil red O staining of HepG2 treated by FFAs and 500 ng/ml AREG for 24 h, respectively. ( H-J ) Macroscopic appearance of liver, liver weight, and liver TG contents of the control and AREG treated mice under HFD condition for 12 weeks ( n = 5 biologically independent mice per condition) (Scale bar, 1 cm). ( K ) Representative image of H&E staining and Oil Red O staining images of the liver sections from the indicated groups ( n = 5 biologically independent mice per condition) (Scale bar, 100 μm). ( L ) Detection of TG levels in HepG2 cells transfected with siRNA targeting EGFR or negative control after exposed to OA/PA and 500 ng/ml AREG for 24 h. ( M ) Western blotting of EGFR in HepG2 cells transfected with siRNA targeting EGFR or negative control and exposed to OA/PA and 500 ng/ml AREG for 24 h ns, not significant; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001 (unpaired two-tailed Student’s t test; mean ± SEM). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The following day, the membrane was incubated with a horseradish peroxidase (HRP)-conjugated secondary antibody for 1 h. The antibodies for immunoblots including KCNMA1 (75-022, L6/60, NeuroMab, Mouse/IgG, 1:1000), AREG (A1860, ABclonal, Rabbit/IgG, 1:1000), Erk (4695, CST, Rabbit/IgG, 1:1000), p-Erk (4370, CST, Rabbit/IgG, 1:1000), EGFR (4267, CST, Rabbit/IgG, 1:1000), GAPDH (2118, CST, Rabbit/IgG, 1:1000) and Tubulin (T0023, Affnity Biosciences, Mouse/IgG, 1:1000).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Knockdown, Staining, Control, Transfection, Negative Control, Western Blot, Two Tailed Test

Journal: Molecular Metabolism

Article Title: Hepatic stellate cell-specific Kcnma1 deletion mitigates metabolic dysfunction-associated steatotic liver disease progression via upregulating Amphiregulin secretion

doi: 10.1016/j.molmet.2025.102164

Figure Lengend Snippet: Downregulation of AREG in the liver of patients with MASLD. ( A ) Representative immunofluorescence results of normal and MASLD patient liver sections, arrows indicate hepatocellular ballooning (Scale bar, 50 μm). ( B ) Immunofluorescence staining for AREG and Lrat in normal and MASLD patient livers (Scale bar, 50 μm). ( C ) Diagrammatic illustration of the suggested mechanism by which Kcnma1 KO HSCs prevent lipid accumulation in hepatocytes by secreting AREG. This graphical work model was generated by applying the BioRender ( https://biorender.com/ ).

Article Snippet: The following day, the membrane was incubated with a horseradish peroxidase (HRP)-conjugated secondary antibody for 1 h. The antibodies for immunoblots including KCNMA1 (75-022, L6/60, NeuroMab, Mouse/IgG, 1:1000), AREG (A1860, ABclonal, Rabbit/IgG, 1:1000), Erk (4695, CST, Rabbit/IgG, 1:1000), p-Erk (4370, CST, Rabbit/IgG, 1:1000), EGFR (4267, CST, Rabbit/IgG, 1:1000), GAPDH (2118, CST, Rabbit/IgG, 1:1000) and Tubulin (T0023, Affnity Biosciences, Mouse/IgG, 1:1000).

Techniques: Immunofluorescence, Staining, Generated

Journal: eLife

Article Title: Atlas of Fshr expression from novel reporter mice

doi: 10.7554/eLife.93413

Figure Lengend Snippet: Frozen sectioned liver tissue was stained for Col1a1 ( A ), CD31 ( B ), or KCNMA1 ( C ), followed by florescent imaging for Fshr-ZsGreen and each of these stained molecules simultaneously. In the section stained for Col1a1 ( A ), two representative areas are shown at magnifications of ×400 and ×1000 as indicated: (1) hepatic cells with a central vein (CV) ( a and c ) and (2) hepatic artery ( b and d ). In the section stained for CD31 ( B ), a representative area of hepatic cells with a CV is shown at lower ( a ) and higher ( b ) magnifications. In the section stained for KCNMA1, three representative areas are presented at two magnifications as follows: (1) hepatic cells with a CV ( a and b ); (2) hepatic cells with peripheral nerve fibers ( c and d ); and (3) small nerve fibers located around a vein ( e and f ).

Article Snippet: Anti-KCNMA1/BK channel Ab , Bioss , bs-4775R , 1:200.

Techniques: Staining, Imaging

Journal: eLife

Article Title: Atlas of Fshr expression from novel reporter mice

doi: 10.7554/eLife.93413

Figure Lengend Snippet: The primary antibodies used for IFs.

Article Snippet: Anti-KCNMA1/BK channel Ab , Bioss , bs-4775R , 1:200.

Techniques: