Journal: Oncotarget

Article Title: Initiation of tumor dormancy by the lymphovascular embolus

doi: 10.18632/oncotarget.28658

Figure Lengend Snippet: Westen blots of mTOR activity with spontaneous spheroidgenesis of Mary-X as measured by pmTOR (Ser2481) and pmTOR (Ser2448) ( A ); of downstream mTOR substrate p-p70-S6K(Thr389) activity in Mary-X spheroids ( B ); of mTOR activity as measured by pmTOR(Ser2448) in induced MCF-7 spheroids ( C ); of downstream mTOR substrates p-p70-S6K(Thr389), p-p70-S6K(Ser371) and p4E-BP1 in induced MCF-7 spheroids; ( D ); of pmTOR(Ser2448) and downstream mTOR substrates p-p70-S6K(Thr389) and p4E-BP1in induced MCF-7 spheroids ( E ).

Article Snippet: We used the following antibodies for Western blot studies: PI3K Ab Sampler Kit (1:1000 dilution, Rabbit, #9655, Cell Signaling Technology (CST)), AMPK and ACC Antibody Sampler Kit (1:1000 dilution, Rabbit, #9957, (CST)), mTOR Substrates Antibody Sampler Kit (1:1000 dilution, Rabbit, #9862, (CST)), E-Cadherin (24E10) (1:1000 dilution, Rabbit mAb #3195, (CST)) and Calpain 2 Large Subunit (M-type) (1:1000 dilution, #3195, (CST)).

Techniques: Activity Assay

Journal: Oncotarget

Article Title: Initiation of tumor dormancy by the lymphovascular embolus

doi: 10.18632/oncotarget.28658

Figure Lengend Snippet: Westen blots of mTOR and AMPK activities and their correlation with levels of calpain 2 and E-cad/NTF1 in spontaneous spheroidgenesis of Mary-X ( A ); of calpain 2 and E-cad/NTF1 and their correlation with the stages of induced spheroidgenesis of E-cadherin positive MCF-7 cells ( B ); of calpain 2 and its correlation with the stages of induced spheroidgenesis of E-cadherin negative MDA-MB-468 cells ( C ). Westen blots of the effects of calpain inhibition on mTOR and AMPK activities in MCF-7 cells grown as monolayers ( D ) and as induced spheroids ( E ).

Article Snippet: We used the following antibodies for Western blot studies: PI3K Ab Sampler Kit (1:1000 dilution, Rabbit, #9655, Cell Signaling Technology (CST)), AMPK and ACC Antibody Sampler Kit (1:1000 dilution, Rabbit, #9957, (CST)), mTOR Substrates Antibody Sampler Kit (1:1000 dilution, Rabbit, #9862, (CST)), E-Cadherin (24E10) (1:1000 dilution, Rabbit mAb #3195, (CST)) and Calpain 2 Large Subunit (M-type) (1:1000 dilution, #3195, (CST)).

Techniques: Inhibition

Journal: Oncotarget

Article Title: Initiation of tumor dormancy by the lymphovascular embolus

doi: 10.18632/oncotarget.28658

Figure Lengend Snippet: Westen blots of the effects of different pathway inhibitors including rapamycin (mTOR), LY294002 (P13K) and U0126 (MAPK) on mTOR activity in Mary-X spontaneous spheroidgenesis ( A ); of the opposite effects of LY294002 (P13K) on mTOR and AMPK activities in both MCF-7 monolayers (left) and induced spheroids (right) ( B ); of P13K activity during induced spheroidgenesis of MCF-7 cells ( C ); of the differential effects of calpain inhibition on P13K activity in MCF-7 monolayers (left) v induced spheroids (right) ( D ). β-Tubulin housekeeping probe was used to normalize for protein loading on divided blots depicted in , as well as . ACTB housekeeping probe was used to normalize for protein loading on divided blots depicted in , as well as , left and right.

Article Snippet: We used the following antibodies for Western blot studies: PI3K Ab Sampler Kit (1:1000 dilution, Rabbit, #9655, Cell Signaling Technology (CST)), AMPK and ACC Antibody Sampler Kit (1:1000 dilution, Rabbit, #9957, (CST)), mTOR Substrates Antibody Sampler Kit (1:1000 dilution, Rabbit, #9862, (CST)), E-Cadherin (24E10) (1:1000 dilution, Rabbit mAb #3195, (CST)) and Calpain 2 Large Subunit (M-type) (1:1000 dilution, #3195, (CST)).

Techniques: Activity Assay, Inhibition

Journal: Oncotarget

Article Title: Initiation of tumor dormancy by the lymphovascular embolus

doi: 10.18632/oncotarget.28658

Figure Lengend Snippet: TMAs ( A ) were subjected to ERAs and SRAs designed to measure true lymphovascular tumor emboli and distinguish them from tumor clumps showing separation artefact from adjacent stroma ( A – C ). Imaging strategy was predicated on SRA’s recognizing podoplanin (D2-40) and CD31 red colorimetric immunoreactivities (B) and the use of ERA’s recognizing epithelial clustering algorithms eg, the Gaussian kernel ( D ), which selectively recognized the clusters within lymphovascular spaces and imaged them ( E ). SRAs showed, compared to their respective non-embolic areas ( F – I ), increased E-cadherin ( J ), decreased Ki-67 ( K ), decreased mTOR (Serine2481) ( L ) and decreased mTOR (Serine2448) ( M ) signal intensities within the emboli of both IBC and non-IBC cases ( N ). For each of these parameters, the graph depicts calculated means ± SD ( N ). Differences of significance are depicted.

Article Snippet: We used the following antibodies for Western blot studies: PI3K Ab Sampler Kit (1:1000 dilution, Rabbit, #9655, Cell Signaling Technology (CST)), AMPK and ACC Antibody Sampler Kit (1:1000 dilution, Rabbit, #9957, (CST)), mTOR Substrates Antibody Sampler Kit (1:1000 dilution, Rabbit, #9862, (CST)), E-Cadherin (24E10) (1:1000 dilution, Rabbit mAb #3195, (CST)) and Calpain 2 Large Subunit (M-type) (1:1000 dilution, #3195, (CST)).

Techniques: Imaging