Journal: Cell reports

Article Title: Microbiome-derived acidity protects against microbial invasion in Drosophila

doi: 10.1016/j.celrep.2024.114087

Figure Lengend Snippet: Number of CFUs of bacteria per fly infected with Ecc15 (A) and EcN (B). Each invasive organism was screened in axenic flies and gnotobiotic flies colonized with Lp, At, and LpAt. Bacterial load was determined at 3 h, 24 h, and 48 h post-feeding infection. Each point represents bacterial load from an individual fly; bars and error bars represent the median and 95% confidence intervals; limit of detection is 2 × 10 2 CFUs/fly. n = 15 flies per treatment per time point over 3 biological replicates. Statistical significance was determined between flies containing different microbiomes by the Mann-Whitney method. p values are represented as follows: * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001.

Article Snippet: Pectobacterium carotovorum pOM1-GFP (Ecc15) (ATCC 25270, pOM1-GFP) , Basset et al., 2003 , N/A.

Techniques: Bacteria, Infection, MANN-WHITNEY

Journal: Cell reports

Article Title: Microbiome-derived acidity protects against microbial invasion in Drosophila

doi: 10.1016/j.celrep.2024.114087

Figure Lengend Snippet: (A) Scheme describing the multi-organism interaction assay procedure. (B) Multi-organism interaction assays display growth effects of gut microbes Lp and At on invasive organisms Pe, Ecc15, and EcN. Microbiome members were grown in perpendicular streaks for 3 days, and invasive organisms were added to the adjacent quadrants and allowed to grow for an additional day. Zones of inhibition are indicated by dashed lines. (C) Co-culture analysis of invasive organisms with microbiome members. Pe, Ecc15, and EcN were grown in mono-culture or in media previously inoculated with Lp, At, or LpAt. Bars and error bars represent the mean concentration of invasive organisms in CFUs/mL ± SEM. N = 3 biological replicates per condition.

Article Snippet: Pectobacterium carotovorum pOM1-GFP (Ecc15) (ATCC 25270, pOM1-GFP) , Basset et al., 2003 , N/A.

Techniques: Inhibition, Co-Culture Assay, Concentration Assay

Journal: Cell reports

Article Title: Microbiome-derived acidity protects against microbial invasion in Drosophila

doi: 10.1016/j.celrep.2024.114087

Figure Lengend Snippet: (A) Multi-organism interaction assay showing the effects of Lp and At on EcN growth on media containing the pH indicator bromophenol blue. After 3 days of growth, a yellow acidic zone appears around Lp, but it tapers off near the At interaction point. When EcN is added, the zone of inhibition overlaps with the acidic region. (B) pH measurement of microbiome mono-cultures and co-cultures reveals a sharp acidification of culture media by Lp. (C) Density of invasive organisms in media adjusted to pH 4.0 with lactic acid with or without 24 h of prior growth with At. Each bar represents mean CFUs/mL ± SEM of 3 biological replicates. (D) Microbial concentration of invasive organisms in media buffered to pH 6.0 with phosphate buffer with or without 24 h of prior growth with Lp NAB1 . Each bar represents mean CFUs/mL ± SEM for 3 biological replicates. (E) Ratio of Ecc15 microbial load in flies infected on buffered vs. standard fly diets, n = 15 flies per treatment per time point over three biological replicates. Statistical significance was determined for flies of each microbiome status between standard and buffered diets using the Mann-Whitney method. p values are represented as follows: NS, p > 0.05, * p ≤ 0.05, and ** p ≤ 0.01. (F) The pH of the copper cell region of the intestine was approximated by feeding axenic and gnotobiotic flies food soaked with 2% bromophenol blue. Guts were dissected and imaged immediately. A yellow/green color in the copper cell region indicates an acidified environment (pH < 4.6).

Article Snippet: Pectobacterium carotovorum pOM1-GFP (Ecc15) (ATCC 25270, pOM1-GFP) , Basset et al., 2003 , N/A.

Techniques: Inhibition, Concentration Assay, Infection, MANN-WHITNEY

Journal: Cell reports

Article Title: Microbiome-derived acidity protects against microbial invasion in Drosophila

doi: 10.1016/j.celrep.2024.114087

Figure Lengend Snippet: (A) Multi-organism interaction assay showing the effects of Lp WCFS1 Lp TF103 ( ΔldhL ldhD::cat ) and At on EcN growth. Neither an acidic zone nor a zone of inhibition appears around Lp TF103 . (B) pH measurement of Lp WCFS1 and Lp TF103 cultures. (C) Analysis of the effect of Lp LDH activity on EcN growth levels during co-culture. EcN was grown in mono-culture or in medium previously inoculated with Lp WCFS1 or Lp TF103 . Bars and error bars represent the mean concentration of invasive organisms in CFUs/mL ± SEM. N = 3 biological replicates per condition. (D) Number of CFUs of Ecc15 per fly. Ecc15 was screened in axenic flies and gnotobiotic flies colonized with Lp WCFS1 or Lp TF103 . Bacterial load was determined at 3 h, 24 h, and 48 h post-feeding infection. Each point represents bacterial load from an individual fly; bars and error bars represent the median and 95% confidence intervals; limit of detection is 2 × 10 2 CFU/fly. n = 15 flies per treatment per time point over 3 biological replicates. Statistical significance was determined between flies containing different microbiomes by the Mann-Whitney method. p values are represented as follows: * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001.

Article Snippet: Pectobacterium carotovorum pOM1-GFP (Ecc15) (ATCC 25270, pOM1-GFP) , Basset et al., 2003 , N/A.

Techniques: Inhibition, Activity Assay, Co-Culture Assay, Concentration Assay, Infection, MANN-WHITNEY

Journal: Cell reports

Article Title: Microbiome-derived acidity protects against microbial invasion in Drosophila

doi: 10.1016/j.celrep.2024.114087

Figure Lengend Snippet: (A) pH analysis of fly food after 3 days of growth of Lp, At, or 1:1 LpAt, with culture medium added as a control. Bars represent the mean pH of three biological replicates ± SEM. (B) pH analysis of fly food 3 days after the addition of 40 male flies of different microbiome statuses (axenic, Lp, At, LpAt). Bars represent the mean pH ± SEM of three biological replicates. (C) Bacterial load of Pe, Ecc15, and EcN present on fly food initially inoculated with microbiome members Lp, At, or 1:1 LpAt, with culture medium as a control. Each bar represents the mean CFUs/g fly food ± SEM for three biological replicates. (D) Pictorial representation of microbe-microbe interactions between microbiome members and gram-negative invasive bacteria on fly food and during consumption by the host.

Article Snippet: Pectobacterium carotovorum pOM1-GFP (Ecc15) (ATCC 25270, pOM1-GFP) , Basset et al., 2003 , N/A.

Techniques: Control, Bacteria

Journal: Cell reports

Article Title: Microbiome-derived acidity protects against microbial invasion in Drosophila

doi: 10.1016/j.celrep.2024.114087

Figure Lengend Snippet:

Article Snippet: Pectobacterium carotovorum pOM1-GFP (Ecc15) (ATCC 25270, pOM1-GFP) , Basset et al., 2003 , N/A.

Techniques:

Journal: Cell reports

Article Title: Microbiome-derived acidity protects against microbial invasion in Drosophila

doi: 10.1016/j.celrep.2024.114087

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Pectobacterium carotovorum pOM1-GFP (Ecc15) (ATCC 25270, pOM1-GFP) , Basset et al., 2003 , N/A.

Techniques: Virus, Recombinant, Lactate Assay, Software

Journal: Cancer Cell International

Article Title: TACR2 is associated with the immune microenvironment and inhibits migration and proliferation via the Wnt/β-catenin signaling pathway in prostate cancer

doi: 10.1186/s12935-021-02126-0

Figure Lengend Snippet: A The expression of TACR2 in all cancers was obtained using the TIMER database. Among them, in kidney chromophobe, kidney renal clear cell carcinoma, and kidney papillary cell carcinoma (KIRP), the expression levels of TACR2 in tumor tissues were lower than that in normal tissues P values were all lower than 0.05, suggesting statistically significant results. B Pair design was performed in TCGA-PRAD to calculate the expression of TACR2 in tumor tissues and normal tissues of the same patient ( P = 1.341e−06). C The expression of TACR2 in tumor tissues and normal tissues of different patients was calculated by randomization in TCGA-PRAD ( P = 1.192e−12). D Expression of TACR2 in the three clinical stages of PRAD ( P = 5.401e−05). E The expression of TACR2 in different Gleason score groups of PRAD ( P = 4.441e−07). F Survival analysis of the group with high expression of TACR2 and the group with low expression of TACR2 showed that the group's survival rate with high expression of TACR2 was higher ( P = 0.0004, HR = 0.5302). G In GSE46602, TACR2 is enriched in the B cell receptor signaling pathway, cancer pathways, the T cell receptor signaling pathway, and the Wnt signaling pathway

Article Snippet: The antibodies included TACR2 (Proteintech, 25270-1-AP), beta-Catenin (Proteintech, 51067-2-AP), GAPDH (Cell Signaling Technology, 5174S), Cyclin D1 (Cell Signaling Technology, 55506S), Lamin B1 (Cell Signaling Technology, 13435S).

Techniques: Expressing

Journal: Cancer Cell International

Article Title: TACR2 is associated with the immune microenvironment and inhibits migration and proliferation via the Wnt/β-catenin signaling pathway in prostate cancer

doi: 10.1186/s12935-021-02126-0

Figure Lengend Snippet: A Panoramic view of immune infiltration in TCGA-PRAD. B Correlation between TACR2 and various immune cells in the immune microenvironment of PRAD. C TACR2 expression was most positively related to resting mast cells (Pearson COR = 0.22; P < 0.001). D TACR2 was most negatively related to M2 macrophages (Pearson COR = − 0.17; P < 0.001)

Article Snippet: The antibodies included TACR2 (Proteintech, 25270-1-AP), beta-Catenin (Proteintech, 51067-2-AP), GAPDH (Cell Signaling Technology, 5174S), Cyclin D1 (Cell Signaling Technology, 55506S), Lamin B1 (Cell Signaling Technology, 13435S).

Techniques: Expressing

Journal: Cancer Cell International

Article Title: TACR2 is associated with the immune microenvironment and inhibits migration and proliferation via the Wnt/β-catenin signaling pathway in prostate cancer

doi: 10.1186/s12935-021-02126-0

Figure Lengend Snippet: Results of WGCNA analysis in TCGA-PRAD. A According to the cutting height = 120,000, 495 samples were included, and the tree diagram of 265 samples was obtained. B We built scale-free co-expression networks. The best soft threshold was 5, R-squared was 0.9. C A hierarchical cluster tree was constructed, with each leaf representing a gene and each branch representing a co-expression module, and a total of eight co-expression modules were generated. D Correlation coefficients between different factors and co-expression modules, where the Brown module (COR = 0.6; P = 4E−41) and grey module (COR = 0.48; P = 1E−24) had a high correlation with TACR2. The correlation between Brown module membership and gene significance for TACR2 was COR = 0.69; P = 9.5e−74

Article Snippet: The antibodies included TACR2 (Proteintech, 25270-1-AP), beta-Catenin (Proteintech, 51067-2-AP), GAPDH (Cell Signaling Technology, 5174S), Cyclin D1 (Cell Signaling Technology, 55506S), Lamin B1 (Cell Signaling Technology, 13435S).

Techniques: Expressing, Construct, Generated

Journal: Cancer Cell International

Article Title: TACR2 is associated with the immune microenvironment and inhibits migration and proliferation via the Wnt/β-catenin signaling pathway in prostate cancer

doi: 10.1186/s12935-021-02126-0

Figure Lengend Snippet: A In TCGA-PRAD, TACR2 was enriched in adherent junctions, regulation of actin cytoskeleton, regulation of autophagy, and the Wnt signaling pathway. B Biology process analysis of TACR2 in TCGA-PRAD. C Correlation between TACR2 and important related genes in the Wnt pathway. TACR2 negatively correlated with FZD6 ( P < 0.001; COR = − 0.25), GSK3B ( P < 0.001; COR = − 0.24) and MAPK8 ( P < 0.001; COR = − 0.26). TACR2 positively correlated with PRKACA ( P < 0.001; COR = 0.30), PRKCA ( P < 0.001; COR = 0.33) and PRKCB ( P < 0.001; COR = 0.40), Wnt2B ( P < 0.001; COR = 0.26), and Wnt5B ( P < 0.001; COR = 0.45)

Article Snippet: The antibodies included TACR2 (Proteintech, 25270-1-AP), beta-Catenin (Proteintech, 51067-2-AP), GAPDH (Cell Signaling Technology, 5174S), Cyclin D1 (Cell Signaling Technology, 55506S), Lamin B1 (Cell Signaling Technology, 13435S).

Techniques:

Journal: Cancer Cell International

Article Title: TACR2 is associated with the immune microenvironment and inhibits migration and proliferation via the Wnt/β-catenin signaling pathway in prostate cancer

doi: 10.1186/s12935-021-02126-0

Figure Lengend Snippet: Expression of TACR2 and β-catenin in prostate cancer and prostate cancer cell lines. A , B The expression of TACR2 protein in prostate cancer tissues (T) and adjacent tissues (N) ( P < 0.01). A , C The expression of β-catenin in prostate cancer tissues and adjacent tissues ( P < 0.01). D , E After transfection with TACR2 overexpression lentiviral vector and corresponding negative control lentiviral vector in DU145, PC3, and LNCaP, protein levels of TACR2, β-catenin and Cyclin D1 were measured using western blot. F , G After isolating nuclear proteins, the nuclear protein levels of β-catenin in TA-OE and NC cells were measured using western blot. The samples derive from the same experiment and those blots were processed in parallel

Article Snippet: The antibodies included TACR2 (Proteintech, 25270-1-AP), beta-Catenin (Proteintech, 51067-2-AP), GAPDH (Cell Signaling Technology, 5174S), Cyclin D1 (Cell Signaling Technology, 55506S), Lamin B1 (Cell Signaling Technology, 13435S).

Techniques: Expressing, Transfection, Over Expression, Plasmid Preparation, Negative Control, Western Blot

Journal: Cancer Cell International

Article Title: TACR2 is associated with the immune microenvironment and inhibits migration and proliferation via the Wnt/β-catenin signaling pathway in prostate cancer

doi: 10.1186/s12935-021-02126-0

Figure Lengend Snippet: Overexpression of TACR2 inhibited cell activity, proliferation, and migration. A The effect of TACR2 overexpression on the proliferation of DU145, PC3, and LNCaP cells were determined using the EdU assay (magnification ×200). B Overexpression of TACR2 suppressed cell viability according to the CCK-8 assay. C Overexpression of TACR2 inhibited cell migration according to the Transwell assay (magnification ×20). D According to the wound-healing assay, overexpression of TACR2 inhibited cell migration in 5637 and T24 cells (magnification ×20)

Article Snippet: The antibodies included TACR2 (Proteintech, 25270-1-AP), beta-Catenin (Proteintech, 51067-2-AP), GAPDH (Cell Signaling Technology, 5174S), Cyclin D1 (Cell Signaling Technology, 55506S), Lamin B1 (Cell Signaling Technology, 13435S).

Techniques: Over Expression, Activity Assay, Migration, EdU Assay, CCK-8 Assay, Transwell Assay, Wound Healing Assay