Fig. 6

PKM2 depletion suppress LPS-induced-β-catenin activation in kidneys and E11 cultured podocytes. A Representative immunoblots (left panel) and bar graph quantitative assessment (right panel) of pβ-Catenin^S33, pβ-Catenin^Y333, β-Catenin and its downstream targets (WT1 and c-Myc), and nephrin in total kidney lysates harvested from C57bl6/J wild type mice 24 h after PBS or LPS injection (n ≥ 6/group). β-Actin was used as a loading control. *p < 0.05, **p < 0.01 indicate a significant difference between PBS- and LPS-injected mice. ^&p < 0.05, ^&&p < 0.01 indicate a significant difference between Ctrl and KO mice. B Representative immunoblots (left panel) and bar graph quantitative assessment (right panel) of pβ-Catenin^Y33, β-Catenin, WT1, and c-Myc in total cell lysates from differentiated M2R and M2KD podocytes treated with LPS for the indicated durations. β-Actin was used as a loading control. Data is representative of at least three independent experiments. *p < 0.05, **p < 0.01 indicate a significant difference between LPS-treated or non-treated cells. ^&p < 0.05, ^&&p < 0.01 indicate a significant difference between M2R and M2KD cells exposed to the same treatment. C Representative immunoblots (left panel) and bar graph quantitative assessment (right panel) of pβ-Catenin^Y333, β-Catenin, GFP, WT1, nephrin, C-Caspase 3, and PKM2 in in total cell lysates from LPS treated or non-treated M2R, M2KD transfected with empty pCS2 + vector (Ctrl), pCS2 plasmid expressing the constitutively active β-catenin mutant (β-CA), or the pLHCX vector expressing the K433E mutant of PKM2. β-Actin was used as a loading control. Data is representative of at least three independent experiments. *p < 0.05, **p < 0.01 indicate a significant difference between LPS-treated or non-treated cells. ^&p < 0.05, ^&&p < 0.01 indicate a significant difference between M2R and M2KD cells exposed to the same treatment. ^#p < 0.05, ^##p < 0.01 indicate a significant difference between the indicated cell overexpressing the constitutively active mutant of β-catenin (β-CA) or the K433E mutant of PKM2 and M2KD cells exposed to the same treatment. D Caspase 3 activity in M2R, M2KD cells in total cell lysates from LPS treated or non-treated M2R, M2KD transfected with empty pCS2 + vector (Ctrl), pCS2 plasmid expressing the constitutively active β-catenin mutant (β-CA) plasmids, or the pLHCX vector expressing the K433E mutant of PKM2. Data is representative of at least three independent experiments. *p < 0.05, **p < 0.01 indicate a significant difference between LPS-treated or non-treated cells. ^&p < 0.05, ^&&p < 0.01 indicate a significant difference between M2R and M2KD cells exposed to the same treatment. ^#p < 0.05, ^##p < 0.01 indicate a significant difference between the indicated cell overexpressing the constitutively active mutant of β-catenin (β-CA) or the K433E mutant of PKM2 and M2KD cells exposed to the same treatment. E Representative images of chromatin condensation in Hoechst-stained M2R and M2KD podocytes transfected with empty pCS2 + vector (Ctrl) or a pCS2 plasmid expressing the constitutively active β-catenin mutant (β-CA). Scale bar: 100 μm. Images are representative of at least three independent experiments