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Localization of <t>Yap1</t> identified using fluorescent immunofluorescence staining . Immunofluorescence staining of mouse proximal tubular epithelial cells highlighting YAP1 localization in relation to tubular injury markers. YAP1 was detected using an Alexa Fluor 555-conjugated antibody ( red ), and the proximal tubular marker LTL was labeled with FITC ( green ). The cell 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.
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Localization of Yap1 identified using fluorescent immunofluorescence staining . Immunofluorescence staining of mouse proximal tubular epithelial cells highlighting YAP1 localization in relation to tubular injury markers. YAP1 was detected using an Alexa Fluor 555-conjugated antibody ( red ), and the proximal tubular marker LTL was labeled with FITC ( green ). The cell 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.

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: Localization of Yap1 identified using fluorescent immunofluorescence staining . Immunofluorescence staining of mouse proximal tubular epithelial cells highlighting YAP1 localization in relation to tubular injury markers. YAP1 was detected using an Alexa Fluor 555-conjugated antibody ( red ), and the proximal tubular marker LTL was labeled with FITC ( green ). The cell 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

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).

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

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.

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