melanoma cancer cell line a375  (ATCC)

Journal: Pharmaceuticals

Article Title: Design, Synthesis, and Bioactivity Assessment of Modified Vemurafenib Analog

doi: 10.3390/ph18081161

Figure Lengend Snippet: Vemurafenib analogs ( 2a – e ) induce apoptosis in A375 cells. Apoptosis was detected by TUNEL assay. A double-staining technique was used. TUNEL staining by using an in situ cell death detection kit (fluorescein) for apoptotic cell nuclei and DAPI (blue) staining for all cell nuclei. A375 cells were treated for 24 h with or without (control) 5 μM of the test compounds. TUNEL-positive cells were visualized as indicated by green fluorescence staining. The images were obtained by EVOS M5000 microscope (Thermo Fisher, Waltham, MA, USA). Representative images from three independent experiments are shown. Magnificance: 20×. Scale bar: 150 μm.

Article Snippet: The human cutaneous melanoma cancer cell line A375, obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA) and human embryonic kidney HEK293T cell line, obtained from Rio de Janeiro Cell Bank (BCRJ, Rio de Janeiro, Brazil), were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma-Aldrich, St. Louis, MO, USA) supplemented with L-glutamine (2 mM), 10% fetal bovine serum (FBS), 50 IU/mL penicillin and 50 μg/mL streptomycin (Sigma-Aldrich, St. Louis, MO, USA).

Techniques: TUNEL Assay, Double Staining, Staining, In Situ, Control, Fluorescence, Microscopy

Journal: Pharmaceuticals

Article Title: Design, Synthesis, and Bioactivity Assessment of Modified Vemurafenib Analog

doi: 10.3390/ph18081161

Figure Lengend Snippet: Morphological appearance of A375 cells treated with vemurafenib and analogs ( 2a – e ) (5 μM) for 24 h. The red arrows indicate apoptotic bodies, showing membrane blebs and fragmented nucleus. The images were obtained by an EVOS M5000 microscope (Thermo Fisher, Waltham, MA, USA). Representative images from three independent experiments are shown. Magnification of 20×.

Article Snippet: The human cutaneous melanoma cancer cell line A375, obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA) and human embryonic kidney HEK293T cell line, obtained from Rio de Janeiro Cell Bank (BCRJ, Rio de Janeiro, Brazil), were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma-Aldrich, St. Louis, MO, USA) supplemented with L-glutamine (2 mM), 10% fetal bovine serum (FBS), 50 IU/mL penicillin and 50 μg/mL streptomycin (Sigma-Aldrich, St. Louis, MO, USA).

Techniques: Membrane, Microscopy

Journal: Pharmaceuticals

Article Title: Design, Synthesis, and Bioactivity Assessment of Modified Vemurafenib Analog

doi: 10.3390/ph18081161

Figure Lengend Snippet: Effects of vemurafenib and analogs ( 2a – e ) on migration of A375 cells. After incubation with medium alone (controls), and the treatment with the compounds test (0.5 μM) for 24 h, cell migration was assayed through cell-based scratch method. ( A ) The images were obtained by EVOS M5000 microscope (Thermo Fisher, Waltham, MA, USA). Representative images from three independent experiments are shown. Magnification of 10×. Scale bar 50 μm. ( B ) The filled areas were analyzed and quantified using ImageJ software (version 1.54p). Statistical analyses were performed with GraphPad Prism 8.02 and included one-way ANOVA followed by Dunnett’s post hoc test. Statistical significance was defined as * p < 0.05 when compared with the control group.

Article Snippet: The human cutaneous melanoma cancer cell line A375, obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA) and human embryonic kidney HEK293T cell line, obtained from Rio de Janeiro Cell Bank (BCRJ, Rio de Janeiro, Brazil), were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma-Aldrich, St. Louis, MO, USA) supplemented with L-glutamine (2 mM), 10% fetal bovine serum (FBS), 50 IU/mL penicillin and 50 μg/mL streptomycin (Sigma-Aldrich, St. Louis, MO, USA).

Techniques: Migration, Incubation, Microscopy, Software, Control

Journal: Pharmaceuticals

Article Title: Design, Synthesis, and Bioactivity Assessment of Modified Vemurafenib Analog

doi: 10.3390/ph18081161

Figure Lengend Snippet: Effects of vemurafenib and analogs ( 2a – e ) on MMP-9 and MMP-2 expression and activity by A375 cells after incubation with medium alone (controls), and the treatment with the test compounds (1 and 5 μm) for 24 h. Gelatinase activity was assayed through zymography method. Representative images from three independent experiments are shown. The protein bands were analyzed using 6.1 software from Bio-Rad.

Article Snippet: The human cutaneous melanoma cancer cell line A375, obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA) and human embryonic kidney HEK293T cell line, obtained from Rio de Janeiro Cell Bank (BCRJ, Rio de Janeiro, Brazil), were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma-Aldrich, St. Louis, MO, USA) supplemented with L-glutamine (2 mM), 10% fetal bovine serum (FBS), 50 IU/mL penicillin and 50 μg/mL streptomycin (Sigma-Aldrich, St. Louis, MO, USA).

Techniques: Expressing, Activity Assay, Incubation, Zymography, Software