Journal: iScience

Article Title: Panx1 deficiency exacerbates GAN diet-induced obesity by destabilizing β-catenin via GSK3β

doi: 10.1016/j.isci.2026.115098

Figure Lengend Snippet: Panx1 increases β-Catenin activity and promotes its nuclear translocation (A) Western blot shows Panx1 overexpression and knockout efficiency (GAPDH as loading control). (B) Glycosylation patterns of exogenous Panx1. (C and D) Total and non-phosphorylated β-catenin levels in whole cell and nuclear fractions (Lamin B1 as loading control). (E) Immunofluorescence of β-catenin nuclear translocation (red; nuclei, blue). Scale bars, 25 μm. (F) qRT-PCR of Wnt/β-catenin downstream gene expression normalized to PPIA. (G) Western blot of Wnt/β-catenin downstream proteins (GAPDH as loading control). Data are mean ± SD, statistics in (C), (D), (F), and (G): ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001, two-sided Student’s t test.

Article Snippet: β-Catenin (D10A8) XP Rabbit mAb , Cell Signaling Technology , Cat# 8480S; RRID: AB_11127855.

Techniques: Activity Assay, Translocation Assay, Western Blot, Over Expression, Knock-Out, Control, Glycoproteomics, Immunofluorescence, Quantitative RT-PCR, Gene Expression

Journal: iScience

Article Title: Panx1 deficiency exacerbates GAN diet-induced obesity by destabilizing β-catenin via GSK3β

doi: 10.1016/j.isci.2026.115098

Figure Lengend Snippet: Panx1 forms complexes with β-Catenin and GSK-3β (A) Western blot of destruction complex proteins in Panx1-OE and Panx1 −/− 3T3-L1 cells (GAPDH as loading control). (B) Co-IP of Panx1 with β-catenin or destruction complex proteins in HEK293T cells expressing Panx1-HA. (C) Co-IP of β-catenin (total or phosphorylated) with GSK-3β in Panx1-OE and Panx1 −/− cells (GAPDH as loading control for input). (D) Western blot showing β-catenin knockdown efficiency in Panx1-OE cells (GAPDH as loading control). (E) Co-IP of Panx1 with GSK-3β after β-catenin knockdown (GAPDH as loading control for input). (F) Predicted 3D structures of Panx1, β-catenin, and GSK-3β complexes showing conformational changes. (G) Ranking scores of binding affinities among Panx1, β-catenin, and GSK-3β. Data are mean ± SD, statistics in (A): ∗ p < 0.05 and ∗∗ p < 0.01, two-sided Student’s t test.

Article Snippet: β-Catenin (D10A8) XP Rabbit mAb , Cell Signaling Technology , Cat# 8480S; RRID: AB_11127855.

Techniques: Western Blot, Control, Co-Immunoprecipitation Assay, Expressing, Knockdown, Binding Assay

Journal: iScience

Article Title: Panx1 deficiency exacerbates GAN diet-induced obesity by destabilizing β-catenin via GSK3β

doi: 10.1016/j.isci.2026.115098

Figure Lengend Snippet: Effect of Panx1 on cell proliferation (A) Cell viability of Panx1-OE and Panx1 −/− cells. (B–D) Flow cytometry analysis of cell cycle distribution (PI staining) with the quantification of G1, S, and G2/M phases. (E) Western blot confirms β-catenin knockdown and effects on downstream proteins in Panx1-OE cells. (F and G) Flow cytometry and quantification of cell cycle distribution after β-catenin knockdown. Each panel represents an independently generated pair of stable lines, and statistical analyses are performed within each matched pair. Data are mean ± SD, statistics in (A), (D), and (G): ∗∗ p < 0.01 and ∗∗∗ p < 0.001, two-sided Student’s t test.

Article Snippet: β-Catenin (D10A8) XP Rabbit mAb , Cell Signaling Technology , Cat# 8480S; RRID: AB_11127855.

Techniques: Flow Cytometry, Staining, Western Blot, Knockdown, Generated

Journal: The Journal of Biological Chemistry

Article Title: PKM2 inhibitor suppresses kidney fibrogenesis by disrupting YAP-TEAD-CCN2 transcriptional signaling following ischemia–reperfusion injury

doi: 10.1016/j.jbc.2025.111029

Figure Lengend Snippet: Immunofluorescence staining of phospho-β-catenin (p-β-catenin, Ser 675) in mouse kidneys was performed to determine the state of phosphorus metabolism . Immunofluorescence staining of p-β-catenin at Ser675 in mouse renal tissue sections. p-β-catenin was detected using an Alexa Fluor 555-conjugated antibody ( red ); the proximal tubular marker LTL was labeled with FITC ( green ), and nuclei were counterstained with DAPI ( blue ) (N = 6). This scale bar represents 100 μm. Data were acquired using a confocal laser scanning microscope. LTL, Lotus tetragonolobus lectin.

Article Snippet: Primary antibodies against the following proteins were used: CCN2 (CTGF) (D8Z8U) (#86641; CST), phosphorylated YAP (Tyr357) (#ab62751; Abcam), YAP1 (#13584-1-AP; Proteintech), PKM2 (#15822-1-AP; Proteintech), Pan-TEAD (D3F7L) (#13295; CST), PKM1 (#15821-1-AP; Proteintech), β-catenin (D10A8) (#8480; CST), lamin B1 (#PM064MS; MBL), GAPDH (D16H11) (#5174; CST), Smad2/3 (D7G7) (#8685; CST), Phospho-Smad3 (Ser423/425) (#GT1207; GTX), and TEAD2 (#21159-1-AP; Proteintech).

Techniques: Immunofluorescence, Staining, Marker, Labeling, Laser-Scanning Microscopy

Journal: The Journal of Biological Chemistry

Article Title: PKM2 inhibitor suppresses kidney fibrogenesis by disrupting YAP-TEAD-CCN2 transcriptional signaling following ischemia–reperfusion injury

doi: 10.1016/j.jbc.2025.111029

Figure Lengend Snippet: Immunofluorescence staining of p-β-catenin (Ser552) in mouse kidneys was performed to determine the state of phosphorus metabolism . Immunofluorescence staining of p-β-catenin at Ser552 in mouse renal tissue sections. p-β-catenin was detected using an Alexa Fluor 555-conjugated antibody ( red ); the proximal tubular marker LTL was labeled with FITC ( green ), and nuclei were counterstained with DAPI ( blue ) (N = 6). This scale bar represents 100 μm. Data were acquired using a confocal laser scanning microscope. LTL, Lotus tetragonolobus lectin.

Article Snippet: Primary antibodies against the following proteins were used: CCN2 (CTGF) (D8Z8U) (#86641; CST), phosphorylated YAP (Tyr357) (#ab62751; Abcam), YAP1 (#13584-1-AP; Proteintech), PKM2 (#15822-1-AP; Proteintech), Pan-TEAD (D3F7L) (#13295; CST), PKM1 (#15821-1-AP; Proteintech), β-catenin (D10A8) (#8480; CST), lamin B1 (#PM064MS; MBL), GAPDH (D16H11) (#5174; CST), Smad2/3 (D7G7) (#8685; CST), Phospho-Smad3 (Ser423/425) (#GT1207; GTX), and TEAD2 (#21159-1-AP; Proteintech).

Techniques: Immunofluorescence, Staining, Marker, Labeling, Laser-Scanning Microscopy

Journal: The Journal of Biological Chemistry

Article Title: PKM2 inhibitor suppresses kidney fibrogenesis by disrupting YAP-TEAD-CCN2 transcriptional signaling following ischemia–reperfusion injury

doi: 10.1016/j.jbc.2025.111029

Figure Lengend Snippet: Western blot analysis of cell fraction protein expression in 3k-treated and siPKM2-transfected HK-2 cells . A and B , Western blot analysis of cytoplasmic and nuclear protein fractions from 3k-treated HK-2 cells to determine β-catenin and YAP1 nuclear translocation. C and D , western blot analysis of cytoplasmic and nuclear protein fractions from siPKM2-transfected HK-2 cells to determine β-catenin and YAP1 nuclear translocation (N = 6).

Article Snippet: Primary antibodies against the following proteins were used: CCN2 (CTGF) (D8Z8U) (#86641; CST), phosphorylated YAP (Tyr357) (#ab62751; Abcam), YAP1 (#13584-1-AP; Proteintech), PKM2 (#15822-1-AP; Proteintech), Pan-TEAD (D3F7L) (#13295; CST), PKM1 (#15821-1-AP; Proteintech), β-catenin (D10A8) (#8480; CST), lamin B1 (#PM064MS; MBL), GAPDH (D16H11) (#5174; CST), Smad2/3 (D7G7) (#8685; CST), Phospho-Smad3 (Ser423/425) (#GT1207; GTX), and TEAD2 (#21159-1-AP; Proteintech).

Techniques: Western Blot, Expressing, Transfection, Translocation Assay

Journal: The Journal of Biological Chemistry

Article Title: PKM2 inhibitor suppresses kidney fibrogenesis by disrupting YAP-TEAD-CCN2 transcriptional signaling following ischemia–reperfusion injury

doi: 10.1016/j.jbc.2025.111029

Figure Lengend Snippet: Western blot analysis of cross-linked protein expression and Co-IP analysis in HK-2 cells . A , western blot analysis of cross-linked proteins extracted from 3k-treated HK-2 cells. B and C , Co-IP analysis of the interaction between YAP1, β-catenin, and PKM2 in HK-2 cells. IP was performed using protein samples from HK-2 cells overexpressing CCN2. Co-IP with an anti-PKM2 antibody confirmed that both β-catenin and YAP1 co-precipitated with PKM2 (N = 6), ( D ) as illustrated in the schematic diagram. CCN2, cellular communication network factor 2.

Article Snippet: Primary antibodies against the following proteins were used: CCN2 (CTGF) (D8Z8U) (#86641; CST), phosphorylated YAP (Tyr357) (#ab62751; Abcam), YAP1 (#13584-1-AP; Proteintech), PKM2 (#15822-1-AP; Proteintech), Pan-TEAD (D3F7L) (#13295; CST), PKM1 (#15821-1-AP; Proteintech), β-catenin (D10A8) (#8480; CST), lamin B1 (#PM064MS; MBL), GAPDH (D16H11) (#5174; CST), Smad2/3 (D7G7) (#8685; CST), Phospho-Smad3 (Ser423/425) (#GT1207; GTX), and TEAD2 (#21159-1-AP; Proteintech).

Techniques: Western Blot, Expressing, Co-Immunoprecipitation Assay