Journal:

Article Title: PCR Detection, Characterization, and Distribution of Virulence Genes in Aeromonas spp.

doi:

Figure Lengend Snippet: Results of PCR and characteristics of the 232-bp amplicons of reference strains

Article Snippet: Lanes: 1 and 15, DNA size marker VIII (Roche Diagnostics AG; catalog no. 1336 045); 2, a PCR-RFLP 2 of A17 (ATCC 7966), a reference strain of HG1 isolated from canned milk from the United States; 3, PCR-RFLP 2 of A307 (ATCC 51108), a reference strain of HG2 isolated from a fish in France; 4, PCR-RFLP 2 of A14 (LMG 13674), a reference strain of HG3 that is an environmental isolate from Germany; 5, PCR-RFLP type 1 of A914 (CDC 9179), a reference strain of HG6 (source unknown); 6, PCR-RFLP 1 of A312 (ATCC 43979), a reference strain of HG7 that is a fish isolate from France; 7, PCR-RFLP 1 of A10 (CCUG 30356), a reference strain of HG8 that is an environmental isolate from Germany; 8, PCR-RFLP 1 of A901 (ATCC 35624), a reference strain of HG10 that is a clinical isolate (sputum) from the United States; 9, PCR-RFLP 1 of A926 (CDC 3136), a reference strain of HG11 that is a water isolate from the United States; 10, PCR-RFLP 3 of A1645 (ATCC 49659), a reference strain of HG13 that is a clinical isolate from the United States; 11, uncut 232-bp amplicon of strain A1645; 12, PCR-RFLP 2 of a wild-type strain (BP 3) of HG1 that is a clinical isolate of HG1 from Bangladesh; 13, PCR-RFLP 2 of a wild-type strain (CBK 45) of HG3 that is a pork meat isolate from Switzerland; 14, PCR-RFLP 1 of a wild-type strain (BE 6) of HG8 that is an environmental isolate from Bangladesh.

Techniques: Fluorescence In Situ Hybridization

Journal:

Article Title: PCR Detection, Characterization, and Distribution of Virulence Genes in Aeromonas spp.

doi:

Figure Lengend Snippet: PCR amplicon of 232 bp and PCR-RFLP types of 232-bp amplicons of different HGs of reference strains of Aeromonas spp. PCR amplicon and PCR-RFLP types of reference and wild-type strains of Aeromonas spp. on 4% Metaphor agarose gel (Bioconcept, Allschwil, Switzerland) stained with ethidium bromide at 0.5 μg/ml (Bio-Rad). Lanes: 1 and 15, DNA size marker VIII (Roche Diagnostics AG; catalog no. 1336 045); 2, a PCR-RFLP 2 of A17 (ATCC 7966), a reference strain of HG1 isolated from canned milk from the United States; 3, PCR-RFLP 2 of A307 (ATCC 51108), a reference strain of HG2 isolated from a fish in France; 4, PCR-RFLP 2 of A14 (LMG 13674), a reference strain of HG3 that is an environmental isolate from Germany; 5, PCR-RFLP type 1 of A914 (CDC 9179), a reference strain of HG6 (source unknown); 6, PCR-RFLP 1 of A312 (ATCC 43979), a reference strain of HG7 that is a fish isolate from France; 7, PCR-RFLP 1 of A10 (CCUG 30356), a reference strain of HG8 that is an environmental isolate from Germany; 8, PCR-RFLP 1 of A901 (ATCC 35624), a reference strain of HG10 that is a clinical isolate (sputum) from the United States; 9, PCR-RFLP 1 of A926 (CDC 3136), a reference strain of HG11 that is a water isolate from the United States; 10, PCR-RFLP 3 of A1645 (ATCC 49659), a reference strain of HG13 that is a clinical isolate from the United States; 11, uncut 232-bp amplicon of strain A1645; 12, PCR-RFLP 2 of a wild-type strain (BP 3) of HG1 that is a clinical isolate of HG1 from Bangladesh; 13, PCR-RFLP 2 of a wild-type strain (CBK 45) of HG3 that is a pork meat isolate from Switzerland; 14, PCR-RFLP 1 of a wild-type strain (BE 6) of HG8 that is an environmental isolate from Bangladesh.

Article Snippet: Lanes: 1 and 15, DNA size marker VIII (Roche Diagnostics AG; catalog no. 1336 045); 2, a PCR-RFLP 2 of A17 (ATCC 7966), a reference strain of HG1 isolated from canned milk from the United States; 3, PCR-RFLP 2 of A307 (ATCC 51108), a reference strain of HG2 isolated from a fish in France; 4, PCR-RFLP 2 of A14 (LMG 13674), a reference strain of HG3 that is an environmental isolate from Germany; 5, PCR-RFLP type 1 of A914 (CDC 9179), a reference strain of HG6 (source unknown); 6, PCR-RFLP 1 of A312 (ATCC 43979), a reference strain of HG7 that is a fish isolate from France; 7, PCR-RFLP 1 of A10 (CCUG 30356), a reference strain of HG8 that is an environmental isolate from Germany; 8, PCR-RFLP 1 of A901 (ATCC 35624), a reference strain of HG10 that is a clinical isolate (sputum) from the United States; 9, PCR-RFLP 1 of A926 (CDC 3136), a reference strain of HG11 that is a water isolate from the United States; 10, PCR-RFLP 3 of A1645 (ATCC 49659), a reference strain of HG13 that is a clinical isolate from the United States; 11, uncut 232-bp amplicon of strain A1645; 12, PCR-RFLP 2 of a wild-type strain (BP 3) of HG1 that is a clinical isolate of HG1 from Bangladesh; 13, PCR-RFLP 2 of a wild-type strain (CBK 45) of HG3 that is a pork meat isolate from Switzerland; 14, PCR-RFLP 1 of a wild-type strain (BE 6) of HG8 that is an environmental isolate from Bangladesh.

Techniques: Amplification, Agarose Gel Electrophoresis, Staining, Marker, Isolation

Journal: Frontiers in Immunology

Article Title: Ginsenoside Rg3 targets glycosylation of PD-L1 to enhance anti-tumor immunity in non-small cell lung cancer

doi: 10.3389/fimmu.2024.1434078

Figure Lengend Snippet: PD-L1 is highly glycosylated in NSCLC cells. (A) Western blot analysis of PD-L1 expression in H1975, H1299, A549 and HCC827 cells. Red asterisk indicated the glycosylated form of PD-L1. (B) H1975 cells were treated with either 5 μg/mL tunicamycin or 20 μM OSMI-4 for 24 h. The expression of PD-L1 was analyzed by western blot. (C) H1975 cells were treated with either 5 μg/mL tunicamycin or 20 μM OSMI-4 for 24 h. Cell viability was determined (n=3). Data were shown as mean ± SD. n.s., not significant.

Article Snippet: The primary antibodies used were as follows: EGFR (Cat No. A23381, 1:1000, Abclonal), Phospho-EGFR-Y1068(Cat No. AP0301, 1:1000, Abclonal), GSK3β (Cat No.9315, 1:1000, Cell Signaling Technology), phospho-GSK3β Ser9 (Cat No.9336, 1:1000, Cell Signaling Technology), PD-L1 (Cat No. A1645, 1:1000, Abclonal) and β-actin (Cat No. AC038, 1:5000, Abclonal).

Techniques: Western Blot, Expressing

Journal: Frontiers in Immunology

Article Title: Ginsenoside Rg3 targets glycosylation of PD-L1 to enhance anti-tumor immunity in non-small cell lung cancer

doi: 10.3389/fimmu.2024.1434078

Figure Lengend Snippet: Ginsenoside Rg3 attenuates the glycosylation of PD-L1 through inhibiting EGFR signaling. (A) Cell viability (n=3). (B) Relative expression of CD274 after 24 h of Rg3 treatments at indicated concentration (n=3). (C) H1975 cells were treated with 10 ng/mL IFN-γ with or without 50 μM Rg3. The percentage of PD-L1 + cells was determined by flow cytometry. (D) Quantification of PD-L1 + cells (n=3). (E–H) H1975 cells were treated with 25 μM or 50 μM Rg3 for 24 h. The expression of p-EGFR, EGFR, p-GSK3β, GSK3β and PD-L1 were determined by western blot. Quantified results were shown. Data were shown as mean ± SD. * p < 0.05, *** p < 0.001 compared to vehicle group. n.s., not significant.

Article Snippet: The primary antibodies used were as follows: EGFR (Cat No. A23381, 1:1000, Abclonal), Phospho-EGFR-Y1068(Cat No. AP0301, 1:1000, Abclonal), GSK3β (Cat No.9315, 1:1000, Cell Signaling Technology), phospho-GSK3β Ser9 (Cat No.9336, 1:1000, Cell Signaling Technology), PD-L1 (Cat No. A1645, 1:1000, Abclonal) and β-actin (Cat No. AC038, 1:5000, Abclonal).

Techniques: Glycoproteomics, Expressing, Concentration Assay, Flow Cytometry, Western Blot

Journal: Frontiers in Immunology

Article Title: Ginsenoside Rg3 targets glycosylation of PD-L1 to enhance anti-tumor immunity in non-small cell lung cancer

doi: 10.3389/fimmu.2024.1434078

Figure Lengend Snippet: Inhibition of PD-L1 glycosylation enhances T cell mediated tumor cell killing in vitro . (A) Schematic illustration of coculture experiment. (B) Percentage of 7-AAD + cells in CD3 - cells (n=3). (C) Histogram of CD25 after coculture and Rg3 treatment. (D) Percentage of CD25 + CD3 + cells in each group (n=3). (E–G) Concentration of IL-2 (E) , IFN-γ (F) and TNF-α (G) in cell culture supernatant (n=3). Data were shown as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001. n.s., not significant.

Article Snippet: The primary antibodies used were as follows: EGFR (Cat No. A23381, 1:1000, Abclonal), Phospho-EGFR-Y1068(Cat No. AP0301, 1:1000, Abclonal), GSK3β (Cat No.9315, 1:1000, Cell Signaling Technology), phospho-GSK3β Ser9 (Cat No.9336, 1:1000, Cell Signaling Technology), PD-L1 (Cat No. A1645, 1:1000, Abclonal) and β-actin (Cat No. AC038, 1:5000, Abclonal).

Techniques: Inhibition, Glycoproteomics, In Vitro, Concentration Assay, Cell Culture

Journal: Frontiers in Immunology

Article Title: Ginsenoside Rg3 targets glycosylation of PD-L1 to enhance anti-tumor immunity in non-small cell lung cancer

doi: 10.3389/fimmu.2024.1434078

Figure Lengend Snippet: Ginsenoside Rg3 attenuated PD-L1 glycosylation and restored anti-tumor immunity. (A) Western blot analysis of PD-L1 in tumor tissues. Red asterisk indicated the glycosylated form of PD-L1. (B) Relative PD-L1 protein expression (n=3). (C–F) Flow cytometry analysis of GzmB + (C, E) and Perforin + (D, F) CD8 + T cells in tumor microenvironment (n=5). (G) Percentage of CD4 + T cells in tumor microenvironment (n=5). (H) Percentage of CD8 + T cells in tumor microenvironment (n=5). Data were shown as mean ± SD. * p < 0.05, ** p < 0.01 compared to model group.

Article Snippet: The primary antibodies used were as follows: EGFR (Cat No. A23381, 1:1000, Abclonal), Phospho-EGFR-Y1068(Cat No. AP0301, 1:1000, Abclonal), GSK3β (Cat No.9315, 1:1000, Cell Signaling Technology), phospho-GSK3β Ser9 (Cat No.9336, 1:1000, Cell Signaling Technology), PD-L1 (Cat No. A1645, 1:1000, Abclonal) and β-actin (Cat No. AC038, 1:5000, Abclonal).

Techniques: Glycoproteomics, Western Blot, Expressing, Flow Cytometry

Journal: bioRxiv

Article Title: Distinct roles of the two SEC scaffold proteins, AFF1 and AFF4, in regulating RNA Pol II transcription elongation

doi: 10.1101/2022.08.09.503397

Figure Lengend Snippet: (A-C) ChIP-qPCR analysis showed the occupancies of elongation factors (AFF1, CDK9, and SPT5) at the FOS gene in AFF4 mutation serum induced HCT116 cells. (D-E) ChIP-qPCR analysis showing that the elongation factors PAF1 and SPT6 occupancy change at the FOS gene in control and AFF4 mutant cells. (F) The protein levels of Ser2P and Ser5P in AFF4 mutation HCT116 cells. α-Tubulin was used as a loading control. (G-H) Metaplot analysis of Ser2P and Ser5P ChIP-seq for the serum induced genes in AFF4 MT cell line. Ser2P and Ser5P density are plotted in a −3 kb and +5 kb window around the genes. (I) Genome browser tracks of Ser2P and Ser5P ChIP-seq at induction genes FOS and EGR1 in control and AFF4 mutation cells in HCT116.

Article Snippet: Ser2P Pol II (ab5095), Ser5P Pol II (ab5131), PAF1 (ab20662) rabbit polyclonal antibody were purchased from Abcam; SPT5 (A9193), SPT6 (A16434), CSTF2 (A8116), CPSF3 (A12368), SYMPK (A8722), Xrn2 (A18350), PPP1CB (A13528), PPP1CC (A4025) rabbit polyclonal antibody were purchased from ABclonal.

Techniques: Mutagenesis, ChIP-sequencing