Journal: Toxins

Article Title: Scorpion Peptide Smp24 Exhibits a Potent Antitumor Effect on Human Lung Cancer Cells by Damaging the Membrane and Cytoskeleton In Vivo and In Vitro

doi: 10.3390/toxins14070438

Figure Lengend Snippet: Viability and morphology changes of cancer cells induced by Smp24. ( A ) Viability of A549, H3122, PC-9, H460 and MRC-5 cells treated with Smp24 for 24 h. ( B ) Proliferation of A549 cells treated with the indicated concentrations of Smp24 for 24 h. Panels ( a – d ) are sequentially the cells treated with 0, 1.25, 2.5, and 5 μM of Smp24. ( C ) Cell morphology changes after treatment with Smp24 for 24 h. Panels ( a – f ) are sequentially the cells treated with 0, 1.25, 2.5, 5, 10 and 20 μM of Smp24. Scale bar, 100 μm. Data are normalized to control and presented as mean ± SEM (n = 3).

Article Snippet: The peptide shows stronger cytotoxicity against leukemic tumor cell lines (KG1-a and CCRF-CEM) than the normal cell lines (CD34 + , HRECs and HACAT) [ ], which indicates that Smp24 may be a potential ACP.

Techniques: Control

Journal: Toxins

Article Title: Scorpion Peptide Smp24 Exhibits a Potent Antitumor Effect on Human Lung Cancer Cells by Damaging the Membrane and Cytoskeleton In Vivo and In Vitro

doi: 10.3390/toxins14070438

Figure Lengend Snippet: Necrosis induced by Smp24 in A549 cells. ( A ) LDH release of A549 cells induced by Smp24 (0–20 µM) for 12, 24 and 48 h. ( B ) SEM analysis of morphological structure of A549 cells. Panel ( a ): control A549 cells; panels ( b , c ): A549 cells treated by 2.5 and 5 µM of Smp24. Scale bar, 10 μm. ( C ) Representative flow cytometry analysis of calcein AM changes in A549 cells treated with Smp24 for 24 h. Panels ( a – d ): cells stained with calcein AM; panels ( a’ – d’ ): cells treated by calcein AM + CoCl 2 ; Panels ( a – a’ , b – b’ , c – c’ , d – d’ ): cells treated by 0, 1.25, 2.5 and 5 µM of Smp24, respectively. Results are presented as mean ± SEM (n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 are considered statistically significant as compared to the control. ( D , E ) Effects of inhibitors on the viability and the LDH release of Smp24-treated A549 cells. A549 cells were pre-incubated with the inhibitors (40 μM Necrostatin-1, 1 μM Cyclosporine A) for 30 min before being further incubated with 5 μM Smp24 for 12 h. ( F ) Fluorescence microscope observation of 5 μM FITC-labeled Smp24 internalized in A549 cells after co-incubation for 24 h. ( a , b ) Control (no treatment) group; ( a’ , b’ ) 24 h treatment group; panels ( c , c’ ) are the merged figure. Yellow arrow: nuclear fragmentation or the apoptotic body. Scale bar, 20 μm.

Article Snippet: The peptide shows stronger cytotoxicity against leukemic tumor cell lines (KG1-a and CCRF-CEM) than the normal cell lines (CD34 + , HRECs and HACAT) [ ], which indicates that Smp24 may be a potential ACP.

Techniques: Control, Flow Cytometry, Staining, Incubation, Fluorescence, Microscopy, Labeling

Journal: Toxins

Article Title: Scorpion Peptide Smp24 Exhibits a Potent Antitumor Effect on Human Lung Cancer Cells by Damaging the Membrane and Cytoskeleton In Vivo and In Vitro

doi: 10.3390/toxins14070438

Figure Lengend Snippet: Effects of Smp24 on the motility and cytoskeleton reorganization of A549 cells. ( A ) Fluorescence staining images of F-actin. Upper panel: cells stained with rhodamine-phalloidin; middle panel: cells stained with DAPI; lower panel: the merging images of cells stained by rhodamine–phalloidin and DAPI. A549 cells were treated with Smp24 (0, 2.5, and 5 μM) for 24 h and successively stained by rhodamine-phalloidin as well as DAPI before fluorescence microscopy observation. White arrow: disordered microfilament bundles. Scale bar, 20 μm. ( B ) Representative pictures of A549 cells in the scratch migration assay at 0 and 24 h following incubation with Smp24 and PBS. Panels ( a – a’ , b – b’ , c – c’ , d – d’ ): the cells treated by 0, 0.3, 0.6 and 1.2 µM of Smp24, respectively. ( C ) Statistical analysis for the scratch migration assay. ( D ) Typical images of A549 cells in the transwell invasion assay following culture with Smp24 and PBS for 24 h. Panels ( a – d ): the cells treated by 0, 0.3, 0.6 and 1.2 µM of Smp24, respectively. ( E ) Statistical analysis of relative mRNA contents of MMP-2, MMP-9, TIMP-1 and TIMP-2. Results are mean ± SEM (n = 3). ** p < 0.01 and *** p < 0.001 are considered statistically significant as compared to the control group without Smp24.

Article Snippet: The peptide shows stronger cytotoxicity against leukemic tumor cell lines (KG1-a and CCRF-CEM) than the normal cell lines (CD34 + , HRECs and HACAT) [ ], which indicates that Smp24 may be a potential ACP.

Techniques: Fluorescence, Staining, Microscopy, Migration, Incubation, Transwell Invasion Assay, Control

Journal: Toxins

Article Title: Scorpion Peptide Smp24 Exhibits a Potent Antitumor Effect on Human Lung Cancer Cells by Damaging the Membrane and Cytoskeleton In Vivo and In Vitro

doi: 10.3390/toxins14070438

Figure Lengend Snippet: In vivo antitumor effects of Smp24. ( A ) Experimental schedule for xenograft mice. ( B ) Images of tumors from sacrificed nude mice. ( C ) Tumor weight. ( D ) Tumor volume. ( E ) Body weight change. ( F ) The weight of heart, liver, spleen, lung and kidney. ( G ) HE staining analysis of tumor tissue. Scale bar, 50 μm. ( H ) Immunohistochemical analysis of cleaved caspase-3 in tumor tissue from lung carcinoma xenografts. Panel ( a ): control group; panel ( b ): Smp24-treated group. Scale bar, 50 μm. Data presented are mean ± SEM (n = 5). ns: no significance, * p < 0.05, ** p < 0.01 and *** p < 0.001 are considered statistically significant as compared to the control group without Smp24 treatment. Yellow arrow: enlarged tumor cells. Blue arrow: shrinking cells. White arrow: positive apoptotic staining (brown areas).

Article Snippet: The peptide shows stronger cytotoxicity against leukemic tumor cell lines (KG1-a and CCRF-CEM) than the normal cell lines (CD34 + , HRECs and HACAT) [ ], which indicates that Smp24 may be a potential ACP.

Techniques: In Vivo, Staining, Immunohistochemical staining, Control

Journal: Toxins

Article Title: Scorpion Peptide Smp24 Exhibits a Potent Antitumor Effect on Human Lung Cancer Cells by Damaging the Membrane and Cytoskeleton In Vivo and In Vitro

doi: 10.3390/toxins14070438

Figure Lengend Snippet: Acute toxicity analysis of Smp24. ( A – D ) The level of ALT, AST, BUN, Cre in serum of normal mice after 48 h treatment with 0, 5 and 10 mg/kg Smp24. ( E – I ) The weight of heart, liver, spleen, lung and kidney of mice after 48 h treatment with 0, 5 and 10 mg/kg Smp24. ns: no significance.

Article Snippet: The peptide shows stronger cytotoxicity against leukemic tumor cell lines (KG1-a and CCRF-CEM) than the normal cell lines (CD34 + , HRECs and HACAT) [ ], which indicates that Smp24 may be a potential ACP.

Techniques:

Journal: Science (New York, N.Y.)

Article Title: Defining the human C2H2 zinc-finger degrome targeted by thalidomide analogs through CRBN

doi: 10.1126/science.aat0572

Figure Lengend Snippet: (A) HEK239T WT and CRBN−/− cells expressing the 11 ZFs identified in the screen in the degradation reporter were treated for 20 hours with drug then analyzed by flow cytometry to measure the DMSO-normalized ratio of eGFP/mCherry fluorescence (exp. rep. = 3, tech. rep. =3, dots represent average of experimental replicates, error bars indicate range). ZFs not shown are in fig S2A. (B) HEK293T WT or CRBN−/− cells expressing full-length (FL), FL with ZF deleted (del ZF), or ZF alone cDNA constructs were treated for 20 hours with DMSO or 1μM drug then analyzed using flow cytometry to measure the DMSO-normalized ratio of eGFP/mCherry fluorescence (exp. rep. = 3, tech. rep. = 3, bar heights indicate mean of exp. rep., error bars indicate 95% CI). Proteins not shown are in fig. S2B. (C) KG1 (human acute myeloid leukemia), (D) WM266–4 (human melanoma), and (E) MOLM-16 cells (human acute myeloid leukemia) were treated with DMSO or 1μM drug for 20 or 24 hours after which protein lysates were harvested, run on a polyacrylamide gel, and immunoblotted for the specified targets (images representative of 3 experimental replicates).

Article Snippet: 2 , KG1 , Acute myeloid leukemia (AML) , Broad Institute Cell Line Repository , RPMI + 10% Fetal bovine serum + penicillin and streptomycin , 37°C , Male , Yes, by Broad cell line repository.

Techniques: Expressing, Flow Cytometry, Fluorescence, Construct

Journal: Science (New York, N.Y.)

Article Title: Defining the human C2H2 zinc-finger degrome targeted by thalidomide analogs through CRBN

doi: 10.1126/science.aat0572

Figure Lengend Snippet: Cell Lines

Article Snippet: 2 , KG1 , Acute myeloid leukemia (AML) , Broad Institute Cell Line Repository , RPMI + 10% Fetal bovine serum + penicillin and streptomycin , 37°C , Male , Yes, by Broad cell line repository.

Techniques: