Journal: Journal of translational medicine

Article Title: Targeting KK-LC-1 inhibits malignant biological behaviors of triple-negative breast cancer.

doi: 10.1186/s12967-023-04030-9

Figure Lengend Snippet: Fig. 4 In vivo experiments presented that KK-LC-1 can regulate the malignant biological behaviors of triple-negative breast cancer. A In vivo tumorigenic models of NC and KD nude mice were successfully established. B Display of tumorigenic specimens in NC KD nude mice. C The volume change of tumor formation in nude mice. D Weight of tumor formation in nude mice. E Detection the expression of PI3K/AKT/mTOR pathway related proteins by immunohistochemistry in NC tumors. F Detection the expression of PI3K/AKT/mTOR pathway related proteins by immunohistochemistry in KD tumors. G Detection the expression of Caspase-3 and PCNA by immunohistochemistry in NC tumors. H Detection the expression of Caspase-3 and PCNA by immunohistochemistry in KD tumors. I Detection the expression of KK-LC-1, MAL2, MUC1 by immunohistochemistry in NC tumors. J Detection the expression of KK-LC-1, MAL2, MUC1 by immunohistochemistry in KD tumors. (**P < 0.01)

Article Snippet: The screening of small-molecule compounds targeting KK-LC-1 was completed by the MCE Company.

Techniques: In Vivo, Expressing, Immunohistochemistry

Journal: Journal of translational medicine

Article Title: Targeting KK-LC-1 inhibits malignant biological behaviors of triple-negative breast cancer.

doi: 10.1186/s12967-023-04030-9

Figure Lengend Snippet: Fig. 6 Validating the correlations between KK-LC-1 protein expression and MAL2 and MUC1 protein expression in triple-negative breast cancer specimens. A Representative pictures of high KK-LC-1 protein expression (Left: ×200 magnification. Right: ×400 magnification). B Representative images of high MAL2 protein expression (Left: ×200 magnification. Right: ×400 magnification). C Scatter plot of the correlation between KK-LC-1 and MAL2 protein expression. D Representative images of high MUC1 protein expression (Left: ×200 magnification. Right: ×400 magnification). E Scatter plot of the correlation between KK-LC-1 and MUC1 protein expression. F The schematic diagram demonstrates that KK-LC-1 regulates the biological behaviors of triple-negative breast cancer cells through the MAL2/MUC1-C/PI3K/AKT/mTOR pathway

Article Snippet: The screening of small-molecule compounds targeting KK-LC-1 was completed by the MCE Company.

Techniques: Expressing

Journal: Journal of translational medicine

Article Title: Targeting KK-LC-1 inhibits malignant biological behaviors of triple-negative breast cancer.

doi: 10.1186/s12967-023-04030-9

Figure Lengend Snippet: Fig. 7 Drug susceptibility test of Z8 small-molecule compound in different breast cancer cell lines. And the inhibitory effect of Z8 on malignant biological behaviors of triple-negative breast cancer cells. A Z8 small-molecule compound molecular structure. B, C Molecular docking assay to analyze the binding ability of Z8 to KK-LC-1 pure protein. D Calculation of EC50 values of Z8 in MDA-MB-231 cells. E Calculation of EC50 values of Z8 in MDA-MB-468 cells. F Analysis of the inhibition rate of MCF7 cell proliferation by Z8 at different concentrations. G Analysis of the inhibition rate of SKBR3 cell proliferation by Z8 at different concentrations. H Analysis of the inhibition rate of MDA-MB-231/KD cell proliferation by Z8 at different concentrations. I Inhibitory effect of Z8 on the cell proliferation. J Inhibitory effect of Z8 on the wound healing ability. K Inhibitory effect of Z8 on the invasive ability. L Inhibitory effect of Z8 on the migration ability. M Z8 promotes apoptosis. N Z8 blocks cell cycle. (**P < 0.01)

Article Snippet: The screening of small-molecule compounds targeting KK-LC-1 was completed by the MCE Company.

Techniques: Docking Assay, Binding Assay, Inhibition, Migration

Journal: Frontiers in Microbiology

Article Title: Enterovirus 71 Antagonizes Antiviral Effects of Type III Interferon and Evades the Clearance of Intestinal Intraepithelial Lymphocytes

doi: 10.3389/fmicb.2021.806084

Figure Lengend Snippet: IFN-λ effectively inhibits EV71 infection in intestinal epithelial cells. (A) HT29 and Jurkat T cells were uninfected or infected with EV71 at an MOI of 1, qRT-PCR was used to detect transcriptional upregulation of both IFN-β, IFN-λ1, and IFN-λ2 at 48 hpi. (B) At the same infection conditions, the production and secretion of IFN-β, IFN-λ1, and IFN-λ2 proteins in the supernatant were detected by ELISA. Mock was cells without infection. (C) HT29 and Jurkat T cells were stimulated without or with 10 ng/ml recombinant protein IFN-β or IFN-λ1 for 12 h. Then the expression levels of ISG54, ISG15, PKR, and OAS were measured by qRT-PCR. Calculated the expression level of each gene relative to the expression of GAPDH and normalized it to mock-treated cells. Control was cells without treatment. (D,E) HT29 cells were pretreated with 10 ng/ml IFN-λ or 10 ng/ml IFN-β for 6 h, respectively, and then infected with EV71 at MOI of 1 for 24 h. The virus replication was monitored by qRT-PCR and plaque experiments. Control was cells with infected but without IFN treatment. Dates were presented as mean ± SD of three replicates ( n = 3 independent experiments, ** P < 0.01, *** P < 0.001, *⁣*⁣** P < 0.0001 and ns, not significant).

Article Snippet: The secretion levels of IFN-β, IFN-λ1, and IFN-λ2 proteins in the supernatant were detected by commercially available enzyme-linked immunosorbent assay kits, including Human IFN-β ELISA Kit (LIANKE, EK1236), Human IL-28A ELISA Kit (BOSTER, EK0969), and Human IL-29 ELISA Kit (BOSTER, EK0964).

Techniques: Infection, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Recombinant, Expressing, Control, Virus

Journal: Frontiers in Microbiology

Article Title: Enterovirus 71 Antagonizes Antiviral Effects of Type III Interferon and Evades the Clearance of Intestinal Intraepithelial Lymphocytes

doi: 10.3389/fmicb.2021.806084

Figure Lengend Snippet: The 2Apro and 3Cpro of EV71 antagonize the antiviral function of IFN-λ and inhibit the expression level of IFN-λ induced by poly(I:C). HT29 cells were transfected with pcDNA3.1-2A (control plasmid: pcDNA3.1) or p-EGFP-3C (control plasmid: p-EGFP). After transfection 24 h, cells were treated with IFN-λ (10 ng/ml) 6 h before infection. Then cells were infected with EV71 at an MOI of 1. Cells and culture mediums were collected at 24 hpi. (A) The protein expression level of VP1 was determined by incubating with anti-EV71 VP1 antibody, followed by staining with Alexa Fluor 555-conjugated secondary antibody. (B) The mRNA expression levels of VP1 were determined by qRT-PCR. (C) Cell culture medium was prepared for virus titer analysis. Control was cells with infected only. IFN-λ group was cells infected and using IFN-λ pretreatment. (D,E) HT29 cells were transfected with pcDNA3.1-2A (pcDNA3.1) or p-EGFP-3C (p-EGFP), and after transfection 24 h, cells were treated with poly(I:C) (4 μg/ml). Cells and culture mediums were collected 24 h after stimulation. The protein expression levels of IFN-λ1 and IFN-λ2 were determined by ELISA, and the mRNA expression levels were determined by qRT-PCR. Control was cells without any treatment. PIC group was cells stimulated only by poly(I:C). Dates were presented as mean ± SD ( n = 3 independent experiments, * P < 0.05, ** P < 0.01 and *** P < 0.001).

Article Snippet: The secretion levels of IFN-β, IFN-λ1, and IFN-λ2 proteins in the supernatant were detected by commercially available enzyme-linked immunosorbent assay kits, including Human IFN-β ELISA Kit (LIANKE, EK1236), Human IL-28A ELISA Kit (BOSTER, EK0969), and Human IL-29 ELISA Kit (BOSTER, EK0964).

Techniques: Expressing, Transfection, Control, Plasmid Preparation, Infection, Staining, Quantitative RT-PCR, Cell Culture, Virus, Enzyme-linked Immunosorbent Assay