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Fig. 3 | Orphanet Journal of Rare Diseases

Fig. 3

From: Synergistic apoptosis following endoplasmic reticulum stress aggravation in mucinous colon cancer

Fig. 3

ERS aggravation by celecoxib plus orlistat combination therapy induces ERS-mediated apoptosis. MTS cell proliferation assay in LS174T cells treated with increasing doses of celecoxib (0–125 μM) a or orlistat (0–125 μM) b or combination of celecoxib and orlistat for 24 h c. Representative pictures of TUNEL assay in LS174T cells treated with single or dual drug therapy for 24 h d. Western blot analysis for ERS and apoptotic markers at 24 h e. f LS174T cells were treated with single or dual drug therapy and stained with JC-1. Diffuse green JC1-monomers indicate mitochondrial depolarization (damage), and punctate red JC1-aggregates indicates intact mitochondrial membrane potential (ΔΨm). g Western blot assay for caspases in LS174T cells following single and dual drug therapy for 24 h. h Representative pictures of TUNEL assay in colonoid cultures from MCC following single and dual drug therapy for 24 h; the bar graph demonstrates mean intensity difference. i Representative pictures from immunofluorescence (IF) analysis of human explant tissue comparing caspase 3 and CHOP between 5 MCC and 5 NMCC following single and dual drug therapy for 24 h; the bar graph demonstrates mean intensity difference. j Western blot assay of apoptotic markers comparing high-MUC2 expressing cells (dnTCF4-LS174T cells exposed to doxycycline for 48 h) versus low-MUC2 expressing cells (dnTCF4-LS174T cells without doxycycline exposure) and comparing stable MUC2 knockdown (KD) in LS174T cells compared to LV control cells following treatment with celecoxib, orlistat, or combination for 24 h. k Western blot assay of CHOP and apoptotic markers comparing stable CHOP knockdown (KD) in LS174T cells compared to LV control cells following treatment with celecoxib, orlistat, or combination for 24 h. Asterisk represents a statistically significant difference compared with the control group (** p < 0.01, *** p < 0.001)

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