Journal: PLOS One

Article Title: Isolation and characterization of a Chlamydia muridarum tc0237 mutant from a genetic screen that is attenuated in epithelial cells

doi: 10.1371/journal.pone.0329637

Figure Lengend Snippet: Equal inoculums of CM TS1 , M7, or M8 were used to infect CMT93, C57, and McCoy cells, and IFU were counted at 24 hpi. The ratio of IFUs that the strains formed in (A) CMT93 versus McCoy cells and in (B) C57 versus McCoy cells were compared. Significance was determined by ordinary one-way ANOVA with Dunnett’s multiple comparisons posttest. Error bars represent standard deviation. **, P < 0.01; ****, P < 0.0001.

Article Snippet: Murine fibroblast (McCoy cells) and murine rectal carcinoma (CMT93) cell lines were obtained from the American Type Culture Collection (ATCC) and were maintained in high glucose Dulbecco’s Modified Eagle medium (DMEM; Cytiva Hyclone) supplemented with 10% fetal bovine serum (FBS; Atlanta Biologicals) non-essential amino acids, and HEPES (DMEM-10).

Techniques: Standard Deviation

Journal: PLOS One

Article Title: Isolation and characterization of a Chlamydia muridarum tc0237 mutant from a genetic screen that is attenuated in epithelial cells

doi: 10.1371/journal.pone.0329637

Figure Lengend Snippet: Equal inocula of CM TS1 , M7, or M8 were used to infect (A) CMT93 and (B) C57 cells + / − IFNγ, and IFU were counted at 24 hpi. IFUs that the strains formed in the cell lines + / − IFNγ were compared. Significance was determined by ordinary one-way ANOVA with Dunnett’s multiple comparison posttest. Error bars represent standard deviation. ***, P < 0.001; ****, P < 0.0001.

Article Snippet: Murine fibroblast (McCoy cells) and murine rectal carcinoma (CMT93) cell lines were obtained from the American Type Culture Collection (ATCC) and were maintained in high glucose Dulbecco’s Modified Eagle medium (DMEM; Cytiva Hyclone) supplemented with 10% fetal bovine serum (FBS; Atlanta Biologicals) non-essential amino acids, and HEPES (DMEM-10).

Techniques: Comparison, Standard Deviation

Journal: PLOS One

Article Title: Isolation and characterization of a Chlamydia muridarum tc0237 mutant from a genetic screen that is attenuated in epithelial cells

doi: 10.1371/journal.pone.0329637

Figure Lengend Snippet: M8, various recombinants, CM TS1 , or CM tsp were used to infect McCoy, ( A ) CMT93, and ( B ) C57 cells at MOIs of 1.0, and inclusions were counted at 24 hpi. Ratios of IFUs that the strains formed in CMT93 or C57 cells divided by the IFUs formed in McCoy cells are shown on the y-axis. Significance was determined by ordinary one-way ANOVA with Tukey’s multiple comparison test. Shared letters indicate groups with p-values greater than 0.05. Groups that do not share a letter have p-values less than 0.05. Error bars represent standard deviation.

Article Snippet: Murine fibroblast (McCoy cells) and murine rectal carcinoma (CMT93) cell lines were obtained from the American Type Culture Collection (ATCC) and were maintained in high glucose Dulbecco’s Modified Eagle medium (DMEM; Cytiva Hyclone) supplemented with 10% fetal bovine serum (FBS; Atlanta Biologicals) non-essential amino acids, and HEPES (DMEM-10).

Techniques: Comparison, Standard Deviation

Journal: PLOS One

Article Title: Isolation and characterization of a Chlamydia muridarum tc0237 mutant from a genetic screen that is attenuated in epithelial cells

doi: 10.1371/journal.pone.0329637

Figure Lengend Snippet: McCoy cells were infected with M8R4-pTC0237-FLAG at an MOI of 1.0 in +/ − aTc and the infected cells were lysed 24 hours later. The lysates, M8, M8R4 237 mut , M8R24 237 wt , CM TS1 , and CM tsp were used to infect fresh McCoy, (A) CMT93, or (B) C57 cells, and IFUs were determined at 24 hpi. (C) McCoy, CMT93, or C57 cells were infected with M8R4-pTC0237-FLAG at MOIs of 0.1, fresh medium + / − aTc was added at 2 hpi, and IFUs were determined at 24 hpi. The number of IFUs formed in each condition is on the y-axis and each dot represents the result of a technical replicate. Significance was determined by ( A-B ) ordinary one-way ANOVA with Tukey’s multiple comparison test or ( C ) unpaired t-test. Shared letters indicate groups with p-values greater than 0.05. Groups that do not share a letter have p-values less than 0.05. Error bars represent standard deviation. ns = not significant.

Article Snippet: Murine fibroblast (McCoy cells) and murine rectal carcinoma (CMT93) cell lines were obtained from the American Type Culture Collection (ATCC) and were maintained in high glucose Dulbecco’s Modified Eagle medium (DMEM; Cytiva Hyclone) supplemented with 10% fetal bovine serum (FBS; Atlanta Biologicals) non-essential amino acids, and HEPES (DMEM-10).

Techniques: Infection, Comparison, Standard Deviation

Journal: PLOS One

Article Title: Isolation and characterization of a Chlamydia muridarum tc0237 mutant from a genetic screen that is attenuated in epithelial cells

doi: 10.1371/journal.pone.0329637

Figure Lengend Snippet: McCoy (black circles) and CMT93 (white squares) cells were infected with Cmu wt , M8, M8R4 237mut , M8R24 237wt , and M8R4-pTC237-FLAG + / − aTc. The number of IFUs produced by these infections was compared to the input inoculum of the primary infection to determine the relative burst size (y-axis) at various hpi (x-axis). Relative burst sizes of infections in McCoy and CMT93 cells were compared by ordinary two-way ANOVA with Bonferroni correction. Graph shows the results from two independent trials, in technical triplicates. Error bars represent standard deviation. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Article Snippet: Murine fibroblast (McCoy cells) and murine rectal carcinoma (CMT93) cell lines were obtained from the American Type Culture Collection (ATCC) and were maintained in high glucose Dulbecco’s Modified Eagle medium (DMEM; Cytiva Hyclone) supplemented with 10% fetal bovine serum (FBS; Atlanta Biologicals) non-essential amino acids, and HEPES (DMEM-10).

Techniques: Infection, Produced, Standard Deviation

Journal: PLOS One

Article Title: Isolation and characterization of a Chlamydia muridarum tc0237 mutant from a genetic screen that is attenuated in epithelial cells

doi: 10.1371/journal.pone.0329637

Figure Lengend Snippet: McCoy (black circles) and CMT93 (white squares) cells were infected with Cmu wt , M8, M8R4 237mut , M8R24 237wt , and M8R4-pTC237-FLAG + / − aTc without centrifugation. The number of IFUs produced was compared to the input IFUs of the primary infection to determine the relative burst size (y-axis) at various hpi (x-axis). Relative burst sizes of infections in McCoy and CMT93 cells were compared by ordinary two-way ANOVA with Bonferroni correction. The graphs show the results from two independent experiment performed in technical triplicates. Error bars indicate standard deviation. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Article Snippet: Murine fibroblast (McCoy cells) and murine rectal carcinoma (CMT93) cell lines were obtained from the American Type Culture Collection (ATCC) and were maintained in high glucose Dulbecco’s Modified Eagle medium (DMEM; Cytiva Hyclone) supplemented with 10% fetal bovine serum (FBS; Atlanta Biologicals) non-essential amino acids, and HEPES (DMEM-10).

Techniques: Infection, Centrifugation, Produced, Standard Deviation

Journal: Gut Microbes

Article Title: Identification of Gut Bacteria such as Lactobacillus johnsonii that Disseminate to Systemic Tissues of Wild Type and MyD88–/– Mice

doi: 10.1080/19490976.2021.2007743

Figure Lengend Snippet: L. johnsonii persist in primary DCs but not macrophages or epithelial cells in vitro. L. johnsonii was cocultured with (a) CMT93 mouse epithelial cells; (b) BMDMs; and (c) BMDCs at MOI of 10:1, for up to 24 h and cell lysates and cell supernatants were plated separately for the indicated time points. Data are from three independent experiments. Statistical analyses were performed by two-tailed Student's t test. p < .05 (denoted by *) was considered statistically significant.

Article Snippet: L. johnsonii strains were cocultured with CMT93 murine intestinal epithelial cell line (ATCC® CCL-223TM), BMDMs (bone-marrow-derived macrophages) and BMDCs (bone-marrow-derived DCs).

Techniques: In Vitro, Two Tailed Test

Journal: Journal of Cellular and Molecular Medicine

Article Title: Mindin serves as a tumour suppressor gene during colon cancer progression through MAPK/ERK signalling pathway in mice

doi: 10.1111/jcmm.15332

Figure Lengend Snippet: Mindin suppresses colon cancer cell proliferation in vitro. A, Analysis of CT26 WT (left side) and CMT93 (right side) cell proliferation in the mindin‐overexpressing cells and control cells by CCK‐8 assay (* P < 0.05). B, Analysis of cell proliferation in the mindin knock‐down cells and control cells by CCK‐8 assay (* P < 0.05). C, Analysis of cell proliferation in the mindin‐overexpressing cells and control cells by BrdU assay (* P < 0.05). D, Analysis of cell proliferation in the mindin knock‐down cells and control cells by BrdU assay (* P < 0.05)

Article Snippet: The CT26 and CMT93 WT cell lines (ATCC, Manassas, VA) were grown in RPMI 1640 and DMEM medium with 10% FBS (Life Technologies, Grand Island, NY) and 1% penicillin G/streptomycin and incubated at 37°C with 95% air and 5% CO 2 .

Techniques: In Vitro, Control, CCK-8 Assay, Knockdown, BrdU Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Mindin serves as a tumour suppressor gene during colon cancer progression through MAPK/ERK signalling pathway in mice

doi: 10.1111/jcmm.15332

Figure Lengend Snippet: Subcutaneous implantation tumour growth of mindin‐overexpressing CMT93 and CT26 WT cells. C57BL/6 and BALB/c mice were subcutaneously injected with stable mindin‐overexpressing CMT93 or CT26 WT cells, or PCMV4 control cells. Tumour size was measured every 3 d for 24 d. A, Images of isolated tumours from the four groups of study mice (n = 5). B, In vivo tumour growth resulting from the mindin‐overexpressing CMT93 or CT26 WT cell (n = 5, * P < 0.05). C, Western blot analysis confirming mindin protein overexpression in the tumour tissues of the four study groups at the end of the study period. Tubulin was used as a protein loading control (n = 5). Upper panel indicates CMT93, and lower panel indicates CT26 WT among A‐C

Article Snippet: The CT26 and CMT93 WT cell lines (ATCC, Manassas, VA) were grown in RPMI 1640 and DMEM medium with 10% FBS (Life Technologies, Grand Island, NY) and 1% penicillin G/streptomycin and incubated at 37°C with 95% air and 5% CO 2 .

Techniques: Injection, Control, Isolation, In Vivo, Western Blot, Over Expression

Journal: Journal of Cellular and Molecular Medicine

Article Title: Mindin serves as a tumour suppressor gene during colon cancer progression through MAPK/ERK signalling pathway in mice

doi: 10.1111/jcmm.15332

Figure Lengend Snippet: Subcutaneous implantation tumour growth of mindin knock‐down CMT93 and CT26 WT cells. C57BL/6 and BALB/c mice were subcutaneously injected with stable mindin knock‐down CMT93 or CT26 WT cells, or PU6 control cells. Tumour size was measured every 3 d for 24 d. A, Images of isolated tumours from the four groups of study mice (n = 5). B, In vivo tumour growth resulting from the mindin knock‐down CMT93 or CT26 WT cell (n = 5, * P < 0.05). C, Western blot analysis confirming mindin protein deficiency in the tumour tissues of the four study groups at the end of the study period. Tubulin was used as a protein loading control (n = 5). Upper panel indicates CMT93, and lower panel indicates CT26 WT among A‐C

Article Snippet: The CT26 and CMT93 WT cell lines (ATCC, Manassas, VA) were grown in RPMI 1640 and DMEM medium with 10% FBS (Life Technologies, Grand Island, NY) and 1% penicillin G/streptomycin and incubated at 37°C with 95% air and 5% CO 2 .

Techniques: Knockdown, Injection, Control, Isolation, In Vivo, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Mindin serves as a tumour suppressor gene during colon cancer progression through MAPK/ERK signalling pathway in mice

doi: 10.1111/jcmm.15332

Figure Lengend Snippet: A, Sequencing chromatograms show the nucleotide mutation of mindin−/− mice using a CRISPR‐Cas system. B, Western blot analysis using antibody against mindin on mice colon tissues. Actin was used as a loading control. C, The tumour images of 21 d after subcutaneous injection of CMT93 colorectal cancer cells in mindin‐knockout and the control mice. Tumour size was measured and quantitatively analysed (n = 5, * P < 0.05). D, Mindin expression was measured by ELISA in the first day and the end point of the model in WT mice serum with or without CRC procedure. E, The cells were isolated from tumour tissues of the AOM/DSS‐induced CRC mice. The procedure of flow cytometry analysis as follows: gated the single cells first, separated cells with the LIVE/DEAD dye and gated the CD45 + cells, then right panels showed the FITC‐CD11b and PE‐F4/80‐stained cells

Article Snippet: The CT26 and CMT93 WT cell lines (ATCC, Manassas, VA) were grown in RPMI 1640 and DMEM medium with 10% FBS (Life Technologies, Grand Island, NY) and 1% penicillin G/streptomycin and incubated at 37°C with 95% air and 5% CO 2 .

Techniques: Sequencing, Mutagenesis, CRISPR, Western Blot, Control, Injection, Knock-Out, Expressing, Enzyme-linked Immunosorbent Assay, Isolation, Flow Cytometry, Staining