Fig. 4: CKMT1 regulated both intrinsic and extrinsic apoptotic pathways in IECs in vitro. | Cell Death & Disease

Fig. 4: CKMT1 regulated both intrinsic and extrinsic apoptotic pathways in IECs in vitro.

From: CKMT1 deficiency contributes to mitochondrial dysfunction and promotes intestinal epithelial cell apoptosis via reverse electron transfer-derived ROS in colitis

Fig. 4

A The protein and B transcriptional expression of CKMT1 in Lovo, HCT116, Caco2, and NCM460 (n = 3). C The CKMT1 knockdown in Lovo were verified via Western blotting. D Confocal microscopy showing the co-localization of CKMT1 and Tomm20 (a marker of mitochondria) protein in IECs. After stimulated with TNF-α (50 ng/ml) and CHX (20 μg/mL) for 4 h, expressions of apoptosis-related proteins were assessed via Western blotting, E in Lovo cells with CKMT1 knockdown (siCK) or control (NC), and F NCM460 cells (n = 4–6). After stimulated with STS (2 μM) for 4 h, apoptosis-related proteins expressions were assessed via western blotting in G Lovo cells, or H NCM460 cells (n = 4). I Apoptosis levels of NCM460 cells after apoptotic stimulation were assessed with Annexin V-FITC/propidium iodide (PI) staining by flow cytometry (n = 3). J Representative images showing drastic change in morphology of intestinal organoid from KOIEC or WT mice, after TNF-α (50 ng/ml) and IFN-γ (50 ng/ml) treatment. K Western blotting showing the expressions of cleaved caspase 3 (n = 4) and E-cadherin (n = 4) in organoids (harvested at 48 h after treatment). *P < 0.05; **P < 0.01; ***P < 0.001.

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