human colonic mucosal epithelial cells  (Procell Inc)

Journal: MedComm

Article Title: Sulfide Quinone Oxidoreductase Alleviates Acute Ulcerative Colitis by Regulating Mitochondrial Dysfunction

doi: 10.1002/mco2.70285

Figure Lengend Snippet: DSS‐induced acute UC significantly reduced SQOR levels. (A) C57BL/6 mice were treated with 3% DSS or water for 7 days, representative immunohistochemical (IHC) staining of SQOR in the colon, red arrows indicated intestinal epithelial cells (scale bar: 200 µm; enlarged scale bar: 100 µm; n = 3). (B, C) Mice were treated with 3% DSS for 7 days, and then colon was prepared to detect SQOR expression using western blot ( n = 4). (D, E) Mice were treated with 3% DSS for 3, 5, and 7 days, representative photograph of colon tissue during DSS‐induced acute UC in colon tissue of WT mice, and the colon length was recorded at days 0, 3, 5, 7 ( n = 3). (F, G) Relative SQOR protein expression in colonic tissues from controls or DSS‐treated WT mice during days 0, 3, 5, 7 using western blot ( n = 3). The data were represented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001. ns, no significant difference.

Article Snippet: NCM460, a normal human colon mucosal epithelial cell line, was purchased from the American Type Culture Collection (ATCC, USA), and maintained in our laboratory.

Techniques: Immunohistochemical staining, Immunohistochemistry, Expressing, Western Blot

Journal: MedComm

Article Title: Sulfide Quinone Oxidoreductase Alleviates Acute Ulcerative Colitis by Regulating Mitochondrial Dysfunction

doi: 10.1002/mco2.70285

Figure Lengend Snippet: SQOR deficiency in the intestinal epithelial cells exacerbates DSS‐induced UC. (A) Western blot analysis of SQOR level in intestinal epithelial cells from Sqor FL/FL and Sqor CKO mice ( n = 3). (B) Immunofluorescence analysis of SQOR (green), Villin (red), and DAPI (blue) in the mouse colonic section was determined by immunofluorescence staining (scale bar: 100 µm; n = 3). (C, D) Daily body weight changes and DAI of Sqor FL/FL and SQOR CKO mice treated with 3% DSS. (E, F) A representative photograph of colon of Sqor FL/FL and Sqor CKO mice on day 7 after DSS treated or not, and the colon length was recorded. (G, H) The histological analysis of colon sections was performed H&E staining from Sqor FL/FL and SQOR CKO mice after DSS treated or not (scale bar: 200 µm; enlarged scale bar: 100 µm), histological scores from Sqor FL/FL and Sqor CKO mice after DSS treated or not ( n = 3). (I, J) Apoptotic cells in colonic sections as determined by TUNEL assay (scale bar: 200 µm; n = 3). (K) Cytokines and chemokines mRNA levels in colon tissues from Sqor FL/FL and Sqor CKO mice after DSS treated or not. The data were represented as mean ± SD. n = 6 mice per group. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. ns, no significant difference.

Article Snippet: NCM460, a normal human colon mucosal epithelial cell line, was purchased from the American Type Culture Collection (ATCC, USA), and maintained in our laboratory.

Techniques: Western Blot, Immunofluorescence, Staining, TUNEL Assay

Journal: MedComm

Article Title: Sulfide Quinone Oxidoreductase Alleviates Acute Ulcerative Colitis by Regulating Mitochondrial Dysfunction

doi: 10.1002/mco2.70285

Figure Lengend Snippet: SQOR deficiency reduces intestinal barrier function in DSS‐induced acute UC in mice. (A) FITC‐dextran of serum determined intestinal permeability from Sqor FL/FL and Sqor CKO mice after DSS treated or not ( n = 4). (B) Representative TEM images of tight junctions between intestinal epithelial cells from Sqor FL/FL and Sqor CKO mice after DSS treatment or not, white arrows indicating tight junctions. (C, D) The occludin in the mouse colon sections was determined by immunofluorescence staining from Sqor FL/FL and Sqor CKO mice after DSS treated or not (scale bar: 200 µm). (E, F) The ZO‐1 in the mouse colon sections was determined by immunofluorescence staining from Sqor FL/FL and Sqor CKO mice after DSS treated or not (scale bar: 200 µm). The data were represented as mean ± SD. n = 3 per group. *p < 0.05, **p < 0.01, ***p < 0.001. ns, no significant difference.

Article Snippet: NCM460, a normal human colon mucosal epithelial cell line, was purchased from the American Type Culture Collection (ATCC, USA), and maintained in our laboratory.

Techniques: Permeability, Immunofluorescence, Staining

Journal: MedComm

Article Title: Sulfide Quinone Oxidoreductase Alleviates Acute Ulcerative Colitis by Regulating Mitochondrial Dysfunction

doi: 10.1002/mco2.70285

Figure Lengend Snippet: SQOR deficiency drives mitochondrial damage in intestinal epithelial cells. (A) Representative mitochondria images of DSS‐stimulated intestinal epithelial cells by TEM from Sqor FL/FL and Sqor CKO mice after DSS treated or not, red arrows indicate damaged mitochondria (scale bar: 5 µm; enlarged scale bar: 500 nm; n = 3). (B) Percentage of damaged mitochondria ( n = 3). (C) Detection of the mRNA levels of Atp5a1, Cox4i1, Uqcrc1, and Ndufab1 in intestinal epithelial cells from Sqor FL/FL and Sqor CKO mice after DSS treated or not ( n = 5). (D) The mtDNA copy number in intestinal epithelial cells from Sqor FL/FL and Sqor CKO mice after DSS treated or not ( n = 6). (E) The ATP level in intestinal epithelial cells from Sqor FL/FL and Sqor CKO mice after DSS treated or not ( n = 4). The data were represented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. ns, no significant difference.

Article Snippet: NCM460, a normal human colon mucosal epithelial cell line, was purchased from the American Type Culture Collection (ATCC, USA), and maintained in our laboratory.

Techniques:

Journal: MedComm

Article Title: Sulfide Quinone Oxidoreductase Alleviates Acute Ulcerative Colitis by Regulating Mitochondrial Dysfunction

doi: 10.1002/mco2.70285

Figure Lengend Snippet: SQOR maintains mitochondrial dynamics homeostasis. (A) Mito‐Tracker was employed to mark mitochondria in NCM460 cells with transfected siRNA‐ SQOR or NC siRNA in the presence or absence of DSS (scale bar: 20 µm; enlarged scale bar: 5 µm). (B) Representative TEM of mitochondria images in intestinal epithelial cells from Sqor FL/FL and Sqor CKO mice after DSS treated or not, red lines represent mitochondrial length measurements. (C) Quantitative analysis of mitochondrial length in TEM images. (D, E) Western blot analysis of SQOR and DRP1 in intestinal epithelial cells from Sqor FL/FL and Sqor CKO mice after DSS treated or not. (F, G) The DRP1 in the mouse colon sections was determined by immunofluorescence staining from Sqor FL/FL and Sqor CKO mice after DSS treated or not (scale bar: 100 µm). The data were represented as mean ± SD. n = 3 per group. *p < 0.05, **p < 0.01. ns, no significant difference.

Article Snippet: NCM460, a normal human colon mucosal epithelial cell line, was purchased from the American Type Culture Collection (ATCC, USA), and maintained in our laboratory.

Techniques: Transfection, Western Blot, Immunofluorescence, Staining

Journal: MedComm

Article Title: Sulfide Quinone Oxidoreductase Alleviates Acute Ulcerative Colitis by Regulating Mitochondrial Dysfunction

doi: 10.1002/mco2.70285

Figure Lengend Snippet: Intestinal epithelial cell function is associated with ROS. (A) Detection of ROS level in intestinal epithelial cells from Sqor FL/FL and Sqor CKO mice after DSS treated or not ( n = 4). (B) GSH/GSSG ratio in intestinal epithelial cells from Sqor FL/FL and Sqor CKO mice after DSS treated or not ( n = 6). (C) The mRNA levels of PGC1α, Nrf1, and Tfam in mouse intestinal epithelial cells from Sqor FL/FL and Sqor CKO mice after DSS treated or not ( n = 6). (D) The mRNA levels of Gpx, Trx2, and Sod2 in mouse intestinal epithelial cells from Sqor FL/FL and Sqor CKO mice after DSS treated or not ( n = 4). (E) The mRNA levels of Ucp2, Ucp4, and Ucp5 in mouse intestinal epithelial cells from Sqor FL/FL and Sqor CKO mice after DSS treated or not ( n = 3). The data were represented as mean ± SD. n = 6 per group. *p < 0.05, **p < 0.01, ***p < 0.001. ns, not significant.

Article Snippet: NCM460, a normal human colon mucosal epithelial cell line, was purchased from the American Type Culture Collection (ATCC, USA), and maintained in our laboratory.

Techniques: Cell Function Assay