Journal: Cell Communication and Signaling : CCS

Article Title: Chemotherapy induced PRL3 expression promotes cancer growth via plasma membrane remodeling and specific alterations of caveolae-associated signaling

doi: 10.1186/s12964-018-0264-8

Figure Lengend Snippet: Induction of PRL3 expression by anticancer drugs in cancer cells. a PRL3 expression is upregulated in B16F0 cells upon treatment with various anticancer drugs. B16F0 cells were treated with 5 μM etoposide, 50 μM cisplatin, 2.5 μM doxorubicin, or 2 μM paclitaxel for 24 h. b , d Dose dependency of PRL3 induction upon doxorubicin and cisplatin administration. B16F0 cells were treated with the indicated amounts of drugs for 24 h. c , e Time dependency of PRL3 induction upon doxorubicin or cisplatin administration. B16F0 cells were treated with 2.5 μM doxorubicin or 50 μM cisplatin. PRL3 expression was analyzed 24 h after treatment or after a second 24 h incubation period without treatment. f PRL3 upregulation upon doxorubicin treatment is a characteristic feature of B16F0, B16F1, and B16F10 cells. The three B16 cell lines were treated with 2.5 μM doxorubicin for 24 h

Article Snippet: B16F0, B16F1, and B16F10 (ATCC, Manassas, Virginia, USA; CRL-6322, CRL-6323, CRL-6475), mouse melanoma cells were maintained in RPMI (Thermo Fischer Scientific, Waltham, Massachusetts, USA); the media was supplemented with 10% FBS (Thermo Fischer Scientific).

Techniques: Expressing, Incubation

Journal: Cell Communication and Signaling : CCS

Article Title: Chemotherapy induced PRL3 expression promotes cancer growth via plasma membrane remodeling and specific alterations of caveolae-associated signaling

doi: 10.1186/s12964-018-0264-8

Figure Lengend Snippet: PRL3 contributes to tumorigenic growth. a PRL3 expression in B16F0 cells artificially expressing (human) PRL3, either in a constitutive or in a conditional (doxycycline-induced) manner. b Growth of B16F0 cells in anchorage-independent conditions. The indicated B16F0 cells were seeded into agar-containing media and cultured for 14 days. Colony formation from single cells was assessed by measuring cell viability with a resazurin-based Alamar blue assay ( n = 3, SEM). * p ≤ 0.005 tested by Student t-test. c Tumor volume changes of subcutaneously injected B16F0 cells in mice. The indicated cells were injected subcutaneously into C57BL/6 J mice ( n = 6, SEM). * p ≤ 0.05 tested by Aspin-Welch t-test. The volumes of the tumors were measured at the indicated time points

Article Snippet: B16F0, B16F1, and B16F10 (ATCC, Manassas, Virginia, USA; CRL-6322, CRL-6323, CRL-6475), mouse melanoma cells were maintained in RPMI (Thermo Fischer Scientific, Waltham, Massachusetts, USA); the media was supplemented with 10% FBS (Thermo Fischer Scientific).

Techniques: Expressing, Cell Culture, Alamar Blue Assay, Injection

Journal: Cell Communication and Signaling : CCS

Article Title: Chemotherapy induced PRL3 expression promotes cancer growth via plasma membrane remodeling and specific alterations of caveolae-associated signaling

doi: 10.1186/s12964-018-0264-8

Figure Lengend Snippet: PRL3 affects the structure of the plasma membrane and localizes to caveolae. In all experiments, PRL3 expression was induced by the addition of doxycycline to the culture media for 24 h. a , b Analysis of PRL3 localization by plasma membrane isolation or by confocal imaging. c GP value changes upon PRL3 expression. Membrane order was assessed by the addition of the membrane incorporating dye di-4-ANNEPDHQ. Data is expressed as the mean of the GP values of at least 80 plasma membrane segments (SEM). * p < 0.001 tested by Student t-test. d PRL3 localizes to the Caveolin 1-rich submembrane fraction in doxycycline-induced B16F0-PRL3 cells e , f Representative TEM images of the membrane of B16F0-PRL3 cells with or without doxycycline induction: e non-induced B16F0-PRL3 cells show an intact caveolae morphology (resolution: 15000X), f PRL3-expressing B16F0 cells show elongated caveolae-like membrane formations (resolution: 15000X). Caveolae-like structures are indicated with arrows

Article Snippet: B16F0, B16F1, and B16F10 (ATCC, Manassas, Virginia, USA; CRL-6322, CRL-6323, CRL-6475), mouse melanoma cells were maintained in RPMI (Thermo Fischer Scientific, Waltham, Massachusetts, USA); the media was supplemented with 10% FBS (Thermo Fischer Scientific).

Techniques: Clinical Proteomics, Membrane, Expressing, Isolation, Imaging

Journal: Cell Communication and Signaling : CCS

Article Title: Chemotherapy induced PRL3 expression promotes cancer growth via plasma membrane remodeling and specific alterations of caveolae-associated signaling

doi: 10.1186/s12964-018-0264-8

Figure Lengend Snippet: PRL3 expression affects integrin ß1 phosphorylation ( a ) Integrin ß1 shows dephosphorylation at Thr788/789 upon the induction of PRL3 expression with doxycycline. ( b ) Raft fractions corresponding to the caveolae region of the membrane from doxycycline-induced B16F0-PRL3 cells were analyzed for the presence of integrin ß1. Only the 150 KDa form of integrin ß1 is present in the caveolae membrane fraction and it cofractionates with PRL3

Article Snippet: B16F0, B16F1, and B16F10 (ATCC, Manassas, Virginia, USA; CRL-6322, CRL-6323, CRL-6475), mouse melanoma cells were maintained in RPMI (Thermo Fischer Scientific, Waltham, Massachusetts, USA); the media was supplemented with 10% FBS (Thermo Fischer Scientific).

Techniques: Expressing, Phospho-proteomics, De-Phosphorylation Assay, Membrane

Journal: Cell Communication and Signaling : CCS

Article Title: Chemotherapy induced PRL3 expression promotes cancer growth via plasma membrane remodeling and specific alterations of caveolae-associated signaling

doi: 10.1186/s12964-018-0264-8

Figure Lengend Snippet: PRL3 expression affects the translocation of Rac1 to the plasma membrane. ( a ) Western blot analysis of isolated total membranes from the indicated cells shows that PRL3 expression results in enhanced translocation of Rac1 to the plasma membrane. ( b ) Representative confocal images of Rac1 plasma membrane localization in PRL3-expressing cells. c Rac1 localizes to the caveolae fraction in the doxycycline-induced B16F0-PRL3 cells. ( d ) Rac1 coimmunoprecipitation with the major caveolae scaffold Caveolin 1 indicates that Rac1 translocate to the caveolae fraction upon PRL3 expression

Article Snippet: B16F0, B16F1, and B16F10 (ATCC, Manassas, Virginia, USA; CRL-6322, CRL-6323, CRL-6475), mouse melanoma cells were maintained in RPMI (Thermo Fischer Scientific, Waltham, Massachusetts, USA); the media was supplemented with 10% FBS (Thermo Fischer Scientific).

Techniques: Expressing, Translocation Assay, Clinical Proteomics, Membrane, Western Blot, Isolation