Journal: Oncotarget

Article Title: Hyperactive ERK and persistent mTOR signaling characterize vemurafenib resistance in papillary thyroid cancer cells

doi: 10.18632/oncotarget.6779

Figure Lengend Snippet: ( A ) Cell lysates from BCPAP cells were collected after 24 hour treatment periods with gradually increasing concentrations of vemurafenib (VMR), subjected to SDS-PAGE and analyzed by Western blot for expression of phospho-ERK1/2 at the end of the 45 day experiment. ( B ) BCPAP and resistant BCPAP cells (from A) were treated with 10 μM VMR for 36 hours and viable cells were counted by trypan blue exclusion assays. **** P <.0001. ( C ) Nthy-ori 3-1 normal thyroid cells, BCPAP, and resistant BCPAP cells were treated with varying concentrations of VMR for 96 hours and then incubated with Alamar blue for 4 hours. Percent absorbance is representative of the difference between measured absorbance at 570 nm and the control absorbance at 600 nm. Data in Figure was published previously in Oncotarget. 2015; 6:39702–13.

Article Snippet: VMR-resistant BCPAP cells were created by intermittently treating BCPAP cells with increasing concentrations of VMR (Chemietek, Indianapolis, IN).

Techniques: SDS Page, Western Blot, Expressing, Incubation, Control

Journal: Oncotarget

Article Title: Hyperactive ERK and persistent mTOR signaling characterize vemurafenib resistance in papillary thyroid cancer cells

doi: 10.18632/oncotarget.6779

Figure Lengend Snippet: ( A–D ) Nthy-ori 3–1, BCPAP, 8505c, and resistant BCPAP cells were subjected to 10 μM VMR for various time points and cytoplasmic extracts were collected and analyzed by Western blot for expression of phospho-ERK1/2, CRAF, phospho-CRAF, and GAPDH (loading control). ( E ) Cytoplasmic extracts from thyroid cell lines were collected after 24 hours untreated and treated with 10 μM VMR and analyzed by Western blot for comparison of CRAF expression. After being subjected to the same 24 hour treatment, thyroid cell lines were collected and lysed using 1% NP-40 buffer. Lysates were subjected to immunoprecipitation using anti-CRAF antibody, SDS-PAGE, and Western blot analysis for CRAF.

Article Snippet: VMR-resistant BCPAP cells were created by intermittently treating BCPAP cells with increasing concentrations of VMR (Chemietek, Indianapolis, IN).

Techniques: Western Blot, Expressing, Control, Comparison, Immunoprecipitation, SDS Page

Journal: Oncotarget

Article Title: Hyperactive ERK and persistent mTOR signaling characterize vemurafenib resistance in papillary thyroid cancer cells

doi: 10.18632/oncotarget.6779

Figure Lengend Snippet: ( A ) Whole cell lysates from thyroid cell lines were collected after 24 hours +/− 10 μM VMR, subjected to SDS-PAGE, and analyzed by Western blot for expression of HER2, HER3, EGFR, and GAPDH (loading control). ( B ) Densitometry of Western blots in A was performed using ImageJ and expressed as percent expression compared to Nthy-ori 3–1 after normalizing each sample to GAPDH. ( C ) Cell lysates were collected as in A and analyzed for expression of phospho-HER3 (Tyr1197) and GAPDH as a loading control. ( D ) Densitometry of Western blots in C was performed using ImageJ and expressed as percent expression compared to untreated for BCPAP and resistant BCPAP samples.

Article Snippet: VMR-resistant BCPAP cells were created by intermittently treating BCPAP cells with increasing concentrations of VMR (Chemietek, Indianapolis, IN).

Techniques: SDS Page, Western Blot, Expressing, Control

Journal: Oncotarget

Article Title: Hyperactive ERK and persistent mTOR signaling characterize vemurafenib resistance in papillary thyroid cancer cells

doi: 10.18632/oncotarget.6779

Figure Lengend Snippet: ( A ) Cell lysates from BCPAP cells were collected after 24 hour treatment periods with gradually increasing concentrations of vemurafenib (VMR), subjected to SDS-PAGE and analyzed by Western blot for expression of phospho-ERK1/2 at the end of the 45 day experiment. ( B ) BCPAP and resistant BCPAP cells (from A) were treated with 10 μM VMR for 36 hours and viable cells were counted by trypan blue exclusion assays. **** P <.0001. ( C ) Nthy-ori 3-1 normal thyroid cells, BCPAP, and resistant BCPAP cells were treated with varying concentrations of VMR for 96 hours and then incubated with Alamar blue for 4 hours. Percent absorbance is representative of the difference between measured absorbance at 570 nm and the control absorbance at 600 nm. Data in Figure was published previously in Oncotarget. 2015; 6:39702–13.

Article Snippet: VMR-resistant BCPAP cells were created by intermittently treating BCPAP cells with increasing concentrations of VMR (Chemietek, Indianapolis, IN).

Techniques: SDS Page, Western Blot, Expressing, Incubation, Control

Journal: Oncotarget

Article Title: Hyperactive ERK and persistent mTOR signaling characterize vemurafenib resistance in papillary thyroid cancer cells

doi: 10.18632/oncotarget.6779

Figure Lengend Snippet: ( A–D ) Nthy-ori 3–1, BCPAP, 8505c, and resistant BCPAP cells were subjected to 10 μM VMR for various time points and cytoplasmic extracts were collected and analyzed by Western blot for expression of phospho-ERK1/2, CRAF, phospho-CRAF, and GAPDH (loading control). ( E ) Cytoplasmic extracts from thyroid cell lines were collected after 24 hours untreated and treated with 10 μM VMR and analyzed by Western blot for comparison of CRAF expression. After being subjected to the same 24 hour treatment, thyroid cell lines were collected and lysed using 1% NP-40 buffer. Lysates were subjected to immunoprecipitation using anti-CRAF antibody, SDS-PAGE, and Western blot analysis for CRAF.

Article Snippet: VMR-resistant BCPAP cells were created by intermittently treating BCPAP cells with increasing concentrations of VMR (Chemietek, Indianapolis, IN).

Techniques: Western Blot, Expressing, Control, Comparison, Immunoprecipitation, SDS Page

Journal: Oncotarget

Article Title: Hyperactive ERK and persistent mTOR signaling characterize vemurafenib resistance in papillary thyroid cancer cells

doi: 10.18632/oncotarget.6779

Figure Lengend Snippet: ( A ) Whole cell lysates from thyroid cell lines were collected after 24 hours +/− 10 μM VMR, subjected to SDS-PAGE, and analyzed by Western blot for expression of HER2, HER3, EGFR, and GAPDH (loading control). ( B ) Densitometry of Western blots in A was performed using ImageJ and expressed as percent expression compared to Nthy-ori 3–1 after normalizing each sample to GAPDH. ( C ) Cell lysates were collected as in A and analyzed for expression of phospho-HER3 (Tyr1197) and GAPDH as a loading control. ( D ) Densitometry of Western blots in C was performed using ImageJ and expressed as percent expression compared to untreated for BCPAP and resistant BCPAP samples.

Article Snippet: VMR-resistant BCPAP cells were created by intermittently treating BCPAP cells with increasing concentrations of VMR (Chemietek, Indianapolis, IN).

Techniques: SDS Page, Western Blot, Expressing, Control