Review



rfi values  (ATCC)


Bioz Verified Symbol ATCC is a verified supplier
Bioz Manufacturer Symbol ATCC manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 93

    Structured Review

    ATCC rfi values
    Rfi Values, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rfi values/product/ATCC
    Average 93 stars, based on 1 article reviews
    rfi values - by Bioz Stars, 2025-02
    93/100 stars

    Images



    Similar Products

    93
    ATCC rfi values
    Rfi Values, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rfi values/product/ATCC
    Average 93 stars, based on 1 article reviews
    rfi values - by Bioz Stars, 2025-02
    93/100 stars
      Buy from Supplier

    86
    Cell Signaling Technology Inc cyclin a2 cst 91500
    Cyclin A2 Cst 91500, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cyclin a2 cst 91500/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    cyclin a2 cst 91500 - by Bioz Stars, 2025-02
    86/100 stars
      Buy from Supplier

    86
    Agilent technologies s zircon 91500
    S Zircon 91500, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/s zircon 91500/product/Agilent technologies
    Average 86 stars, based on 1 article reviews
    s zircon 91500 - by Bioz Stars, 2025-02
    86/100 stars
      Buy from Supplier

    86
    Penglai Nuokang Pharmaceutical Co Ltd zircon rms penglai 13 91500 8 9 plešovice 10 11 qinghu12
    Zircon Rms Penglai 13 91500 8 9 Plešovice 10 11 Qinghu12, supplied by Penglai Nuokang Pharmaceutical Co Ltd, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/zircon rms penglai 13 91500 8 9 plešovice 10 11 qinghu12/product/Penglai Nuokang Pharmaceutical Co Ltd
    Average 86 stars, based on 1 article reviews
    zircon rms penglai 13 91500 8 9 plešovice 10 11 qinghu12 - by Bioz Stars, 2025-02
    86/100 stars
      Buy from Supplier

    86
    Penglai Nuokang Pharmaceutical Co Ltd including gj 1 91500 plešovice mud tank
    Including Gj 1 91500 Plešovice Mud Tank, supplied by Penglai Nuokang Pharmaceutical Co Ltd, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/including gj 1 91500 plešovice mud tank/product/Penglai Nuokang Pharmaceutical Co Ltd
    Average 86 stars, based on 1 article reviews
    including gj 1 91500 plešovice mud tank - by Bioz Stars, 2025-02
    86/100 stars
      Buy from Supplier

    86
    Agdia Inc sra 91500
    Sra 91500, supplied by Agdia Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sra 91500/product/Agdia Inc
    Average 86 stars, based on 1 article reviews
    sra 91500 - by Bioz Stars, 2025-02
    86/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc cyclin a2
    c-MYC expression drives cell cycle progression. (A) Tet-On c-MYC–expressing PDX220 or Panc 10.05 cells treated with vehicle or 2 μg/ml Dox for 3, 5, or 7 d were assessed for cell cycle. Bar graphs show representative data from two independent experiments for PDX220 and one experiment for Panc 10.05. (B–D) c-MYC T58A reactivates the cell cycle in the face of T/CQ co-treatment, but not P/CQ co-treatment. Percentage of cells in each cell cycle phase in (B) Panc 10.05 ( n = 1 from one independent experiment), (C) HPAF-II ( n = 3 from one experiment), or (D) PDX220 cells ( n = 1 from three independent experiments) expressing Tet-On c-MYC T58A are depicted. Cells were treated for 7 d with DMSO (Ctrl), 2 μg/ml Dox, 10–100 nM trametinib and 10–20 μM CQ (T/CQ), D/T/CQ, 2.5 μM palbociclib and 10–20 μM CQ (P/CQ) or D/P/CQ and cell cycle phase was quantitated. Bars represent mean percent cell cycle phase ± SEM. *, P < 0.05; **, P < 0.05; ns, not significant by Student’s two-sided t test. (E) CQ co-treatment does not significantly affect cell cycle status in trametinib- or palbociclib-treated PDAC cells. PDX220 Tet-On c-MYC cells were treated with vehicle control (Ctrl), CQ (20 μM), trametinib (T, 100 nM), palbociclib (P, 2.5 μM), T/CQ, or P/CQ for 7 d and processed for cell cycle analysis ( n = 1 from two independent experiments, mean ± SEM). ns, not significant by Student’s two-tailed t test. (F) Panc 10.05 cells expressing Tet-On c-MYC were treated for 7 d with DMSO (Ctrl), CQ (20 μM), trametinib (T, 100 nM), or palbociclib (P, 2.5 μM) or the combinations (T/CQ and P/CQ), and with or without Dox (D, 2 μg/ml; n = 1). (G) Cell cycle profiles were assessed in MIA-PaCa2 cells after treatment with palbociclib with and without CQ co-treatment. A Student’s two-tailed t test was utilized in order to determine statistical significance of G 0 /G 1 phase between the DMSO control and the experimental groups as well as between the experimental groups in one experiment ( n = 3). ns, not significant; ***, P < 0.001. (H–J) c-MYC T58A expression elevates pRB, cyclinB1, and cyclinA2 expression in the face of trametinib and CQ, but not P/CQ co-treatment. Representative immunoblot analysis of Tet-On c-MYC T58A expressing Panc 10.05 (H), HPAF-II (I), and PDX220 (J) cells that were treated as in B–D (from three independent experiments). Lysates were blotted for c-MYC, pERK1/2, ERK1/2, pRB, RB, E2F1, CCNB1, <t>CCNA2,</t> CCND2, CCNE1, CDK6, p27 KIP1 , p21 CIP1 , and ACTB. (K and L) c-MYC WT elevates pRB, cyclinB1, and cyclinA2 expression in the face of T/CQ, but not P/CQ. Representative immunoblot analysis of Tet-On c-MYC HPAF-II and PDX220 cells that were treated as above (from two independent experiments for HPAF-II and n = 1 experiment for PDX220). Lysates were blotted for c-MYC, pERK1/2, ERK1/2, pRB, RB, E2F1, CCNB1, CCNA2, CCND2, CCNE1, CDK2, CDK4, CDK6, p27 KIP1 , p21 CIP1 , and ACTB. Source data are available for this figure: .
    Cyclin A2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cyclin a2/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    cyclin a2 - by Bioz Stars, 2025-02
    94/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc antibodies against cyclin a2
    The qPCR primers of different genes
    Antibodies Against Cyclin A2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/antibodies against cyclin a2/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    antibodies against cyclin a2 - by Bioz Stars, 2025-02
    94/100 stars
      Buy from Supplier

    86
    National Institute of Standards and Technology standard zircon 91500
    The qPCR primers of different genes
    Standard Zircon 91500, supplied by National Institute of Standards and Technology, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/standard zircon 91500/product/National Institute of Standards and Technology
    Average 86 stars, based on 1 article reviews
    standard zircon 91500 - by Bioz Stars, 2025-02
    86/100 stars
      Buy from Supplier

    Image Search Results


    c-MYC expression drives cell cycle progression. (A) Tet-On c-MYC–expressing PDX220 or Panc 10.05 cells treated with vehicle or 2 μg/ml Dox for 3, 5, or 7 d were assessed for cell cycle. Bar graphs show representative data from two independent experiments for PDX220 and one experiment for Panc 10.05. (B–D) c-MYC T58A reactivates the cell cycle in the face of T/CQ co-treatment, but not P/CQ co-treatment. Percentage of cells in each cell cycle phase in (B) Panc 10.05 ( n = 1 from one independent experiment), (C) HPAF-II ( n = 3 from one experiment), or (D) PDX220 cells ( n = 1 from three independent experiments) expressing Tet-On c-MYC T58A are depicted. Cells were treated for 7 d with DMSO (Ctrl), 2 μg/ml Dox, 10–100 nM trametinib and 10–20 μM CQ (T/CQ), D/T/CQ, 2.5 μM palbociclib and 10–20 μM CQ (P/CQ) or D/P/CQ and cell cycle phase was quantitated. Bars represent mean percent cell cycle phase ± SEM. *, P < 0.05; **, P < 0.05; ns, not significant by Student’s two-sided t test. (E) CQ co-treatment does not significantly affect cell cycle status in trametinib- or palbociclib-treated PDAC cells. PDX220 Tet-On c-MYC cells were treated with vehicle control (Ctrl), CQ (20 μM), trametinib (T, 100 nM), palbociclib (P, 2.5 μM), T/CQ, or P/CQ for 7 d and processed for cell cycle analysis ( n = 1 from two independent experiments, mean ± SEM). ns, not significant by Student’s two-tailed t test. (F) Panc 10.05 cells expressing Tet-On c-MYC were treated for 7 d with DMSO (Ctrl), CQ (20 μM), trametinib (T, 100 nM), or palbociclib (P, 2.5 μM) or the combinations (T/CQ and P/CQ), and with or without Dox (D, 2 μg/ml; n = 1). (G) Cell cycle profiles were assessed in MIA-PaCa2 cells after treatment with palbociclib with and without CQ co-treatment. A Student’s two-tailed t test was utilized in order to determine statistical significance of G 0 /G 1 phase between the DMSO control and the experimental groups as well as between the experimental groups in one experiment ( n = 3). ns, not significant; ***, P < 0.001. (H–J) c-MYC T58A expression elevates pRB, cyclinB1, and cyclinA2 expression in the face of trametinib and CQ, but not P/CQ co-treatment. Representative immunoblot analysis of Tet-On c-MYC T58A expressing Panc 10.05 (H), HPAF-II (I), and PDX220 (J) cells that were treated as in B–D (from three independent experiments). Lysates were blotted for c-MYC, pERK1/2, ERK1/2, pRB, RB, E2F1, CCNB1, CCNA2, CCND2, CCNE1, CDK6, p27 KIP1 , p21 CIP1 , and ACTB. (K and L) c-MYC WT elevates pRB, cyclinB1, and cyclinA2 expression in the face of T/CQ, but not P/CQ. Representative immunoblot analysis of Tet-On c-MYC HPAF-II and PDX220 cells that were treated as above (from two independent experiments for HPAF-II and n = 1 experiment for PDX220). Lysates were blotted for c-MYC, pERK1/2, ERK1/2, pRB, RB, E2F1, CCNB1, CCNA2, CCND2, CCNE1, CDK2, CDK4, CDK6, p27 KIP1 , p21 CIP1 , and ACTB. Source data are available for this figure: .

    Journal: The Journal of Experimental Medicine

    Article Title: MYC-mediated resistance to trametinib and HCQ in PDAC is overcome by CDK4/6 and lysosomal inhibition

    doi: 10.1084/jem.20221524

    Figure Lengend Snippet: c-MYC expression drives cell cycle progression. (A) Tet-On c-MYC–expressing PDX220 or Panc 10.05 cells treated with vehicle or 2 μg/ml Dox for 3, 5, or 7 d were assessed for cell cycle. Bar graphs show representative data from two independent experiments for PDX220 and one experiment for Panc 10.05. (B–D) c-MYC T58A reactivates the cell cycle in the face of T/CQ co-treatment, but not P/CQ co-treatment. Percentage of cells in each cell cycle phase in (B) Panc 10.05 ( n = 1 from one independent experiment), (C) HPAF-II ( n = 3 from one experiment), or (D) PDX220 cells ( n = 1 from three independent experiments) expressing Tet-On c-MYC T58A are depicted. Cells were treated for 7 d with DMSO (Ctrl), 2 μg/ml Dox, 10–100 nM trametinib and 10–20 μM CQ (T/CQ), D/T/CQ, 2.5 μM palbociclib and 10–20 μM CQ (P/CQ) or D/P/CQ and cell cycle phase was quantitated. Bars represent mean percent cell cycle phase ± SEM. *, P < 0.05; **, P < 0.05; ns, not significant by Student’s two-sided t test. (E) CQ co-treatment does not significantly affect cell cycle status in trametinib- or palbociclib-treated PDAC cells. PDX220 Tet-On c-MYC cells were treated with vehicle control (Ctrl), CQ (20 μM), trametinib (T, 100 nM), palbociclib (P, 2.5 μM), T/CQ, or P/CQ for 7 d and processed for cell cycle analysis ( n = 1 from two independent experiments, mean ± SEM). ns, not significant by Student’s two-tailed t test. (F) Panc 10.05 cells expressing Tet-On c-MYC were treated for 7 d with DMSO (Ctrl), CQ (20 μM), trametinib (T, 100 nM), or palbociclib (P, 2.5 μM) or the combinations (T/CQ and P/CQ), and with or without Dox (D, 2 μg/ml; n = 1). (G) Cell cycle profiles were assessed in MIA-PaCa2 cells after treatment with palbociclib with and without CQ co-treatment. A Student’s two-tailed t test was utilized in order to determine statistical significance of G 0 /G 1 phase between the DMSO control and the experimental groups as well as between the experimental groups in one experiment ( n = 3). ns, not significant; ***, P < 0.001. (H–J) c-MYC T58A expression elevates pRB, cyclinB1, and cyclinA2 expression in the face of trametinib and CQ, but not P/CQ co-treatment. Representative immunoblot analysis of Tet-On c-MYC T58A expressing Panc 10.05 (H), HPAF-II (I), and PDX220 (J) cells that were treated as in B–D (from three independent experiments). Lysates were blotted for c-MYC, pERK1/2, ERK1/2, pRB, RB, E2F1, CCNB1, CCNA2, CCND2, CCNE1, CDK6, p27 KIP1 , p21 CIP1 , and ACTB. (K and L) c-MYC WT elevates pRB, cyclinB1, and cyclinA2 expression in the face of T/CQ, but not P/CQ. Representative immunoblot analysis of Tet-On c-MYC HPAF-II and PDX220 cells that were treated as above (from two independent experiments for HPAF-II and n = 1 experiment for PDX220). Lysates were blotted for c-MYC, pERK1/2, ERK1/2, pRB, RB, E2F1, CCNB1, CCNA2, CCND2, CCNE1, CDK2, CDK4, CDK6, p27 KIP1 , p21 CIP1 , and ACTB. Source data are available for this figure: .

    Article Snippet: Antibodies used are as follows (used at 1:1,000 unless noted otherwise): 1:2,000 phospho-T202/Y204 p44/42 MAPK (ERK1/2; CST:4370), total ERK1/2 (CST:4696); p-RB 807/811 1:1,000 (CST 8516S); RB (CST:9309S); c-MYC (CST:5605S); E2F1 (CST:3742S); Cyclin B1 (CST:12231S); Cyclin A2 (CST:91500S); Cyclin D2 (CST:3741S); Cyclin E1 (CST:20808S); CDK2 (CST:2546S); CDK4 (CST:12790S); CDK6 (CST:3136S); p27 KIP1 , (CST:3686S); p21 WAF1/CIP1 (CST:2947S); 1:1,000 Vinculin (CST:13901); 1:2,000 GAPDH (CST:97166S); 1:5,000 β-actin (CST:4970S).

    Techniques: Expressing, Cell Cycle Assay, Two Tailed Test, Western Blot

    Elevated c-MYC expression prevents T/CQ-mediated cell cycle arrest. (A) Representative cell cycle histograms with percentages of cells in G 0 /G 1 , S, and G 2 /M phases are shown for Panc 10.05, HPAF-II, and PDX220 cells expressing Tet-On c-MYC. Cells were treated for 7 d with DMSO (Ctrl), 2 μg/ml Dox, trametinib (T, at 5–100 nM) and CQ (at 5–20 μM; T/CQ), or D/T/CQ. (B) Average percentage of cells in S phase for Tet-On c-MYC–expressing Panc 10.05, HPAF-II, and PDX220 cells that were treated as above ( n = 3 from two independent experiments for each cell type, mean ± SEM). *, P < 0.05; **, P < 0.01; Student’s two-sided t test. (C) Immunoblot analysis of lysates generated from Tet-On c-MYC–expressing cells that were treated as above ( n = 1 for each cell type). Lysates were blotted for c-MYC, pERK1/2 (phospho-T202/Y204), ERK1/2, pRB (phospho-S807/S811), RB, E2F1, CCNB1, CCNA2, CCND2, CCNE1, CDK2, CDK4, CDK6, p27 KIP1 , p21 CIP1 , and ACTB. Densitometry quantitation of c-MYC and pRB protein levels normalized to actin are shown. Source data are available for this figure: .

    Journal: The Journal of Experimental Medicine

    Article Title: MYC-mediated resistance to trametinib and HCQ in PDAC is overcome by CDK4/6 and lysosomal inhibition

    doi: 10.1084/jem.20221524

    Figure Lengend Snippet: Elevated c-MYC expression prevents T/CQ-mediated cell cycle arrest. (A) Representative cell cycle histograms with percentages of cells in G 0 /G 1 , S, and G 2 /M phases are shown for Panc 10.05, HPAF-II, and PDX220 cells expressing Tet-On c-MYC. Cells were treated for 7 d with DMSO (Ctrl), 2 μg/ml Dox, trametinib (T, at 5–100 nM) and CQ (at 5–20 μM; T/CQ), or D/T/CQ. (B) Average percentage of cells in S phase for Tet-On c-MYC–expressing Panc 10.05, HPAF-II, and PDX220 cells that were treated as above ( n = 3 from two independent experiments for each cell type, mean ± SEM). *, P < 0.05; **, P < 0.01; Student’s two-sided t test. (C) Immunoblot analysis of lysates generated from Tet-On c-MYC–expressing cells that were treated as above ( n = 1 for each cell type). Lysates were blotted for c-MYC, pERK1/2 (phospho-T202/Y204), ERK1/2, pRB (phospho-S807/S811), RB, E2F1, CCNB1, CCNA2, CCND2, CCNE1, CDK2, CDK4, CDK6, p27 KIP1 , p21 CIP1 , and ACTB. Densitometry quantitation of c-MYC and pRB protein levels normalized to actin are shown. Source data are available for this figure: .

    Article Snippet: Antibodies used are as follows (used at 1:1,000 unless noted otherwise): 1:2,000 phospho-T202/Y204 p44/42 MAPK (ERK1/2; CST:4370), total ERK1/2 (CST:4696); p-RB 807/811 1:1,000 (CST 8516S); RB (CST:9309S); c-MYC (CST:5605S); E2F1 (CST:3742S); Cyclin B1 (CST:12231S); Cyclin A2 (CST:91500S); Cyclin D2 (CST:3741S); Cyclin E1 (CST:20808S); CDK2 (CST:2546S); CDK4 (CST:12790S); CDK6 (CST:3136S); p27 KIP1 , (CST:3686S); p21 WAF1/CIP1 (CST:2947S); 1:1,000 Vinculin (CST:13901); 1:2,000 GAPDH (CST:97166S); 1:5,000 β-actin (CST:4970S).

    Techniques: Expressing, Western Blot, Generated, Quantitation Assay

    Elevated c-MYC expression does not prevent P/CQ-mediated cell cycle arrest. (A) Panc 10.05, PDX220, and MIA-PaCa2 cells were imaged to assess confluence over time using the IncuCyte Live-Cell Analysis System after treatment with palbociclib (P), CQ, or the combination of both agents (P/CQ). Data represent mean values, and error bars reflect SD of one experiment ( n = 3). One-way ANOVA was conducted to determine statistical difference between the single agent controls (CDK4/6 inhibitor or CQ) and the combination group. *, P < 0.05; **, P < 0.01; ***, P < 0.001. (B) MIA-PaCa2 cells were treated with ribociclib (Ribo) or abemaciclib (Abema) alone or in combination with CQ and confluence was measured over time. A one-way ANOVA was utilized to determine statistical significance between the single agent controls and the combination group. The Ribo/CQ experiment was conducted twice ( n = 2) and the Abema/CQ experiment was conducted once ( n = 3). *, P < 0.05; **, P < 0.01; ***, P < 0.001. (C) Representative images of MIA-PaCa2 cells shown by phase contrast (top) and the corresponding cell mask utilized to estimate well confluence (bottom) for each treatment at 48 h. Bars are 300 μm. (D) Average percentage of cells in S phase for Tet-On c-MYC–expressing Panc 10.05, HPAF-II, and PDX220 cells that were treated for 7 d with DMSO (Ctrl); 2 μg/ml Dox; 5–10 μM CQ and 2.5 μM palbociclib (P/CQ); or D/P/CQ. Control and Dox-treated sample percentages are continued from ; experiments were performed at the same time as P/CQ- and D/P/CQ-treated samples ( n = 3 from two independent experiments, mean ± SEM). *, P < 0.05; **, P < 0.01; ns, not significant by Student’s two-sided t test. (E) Immunoblot analysis of lysates generated from Tet-On c-MYC–expressing cells that were treated for 7 d with DMSO (Ctrl), 2 μg/ml Dox (D), 5–10 μM CQ, 2.5 μM palbociclib (P), or the indicated combinations (representative blots from two independent experiments). Lysates were blotted for c-MYC, pRB (phospho-S807/811), RB, CCNB1, CCNA2, and ACTB. Densitometry quantitation of c-MYC and pRB protein levels normalized to actin are shown. Source data are available for this figure: .

    Journal: The Journal of Experimental Medicine

    Article Title: MYC-mediated resistance to trametinib and HCQ in PDAC is overcome by CDK4/6 and lysosomal inhibition

    doi: 10.1084/jem.20221524

    Figure Lengend Snippet: Elevated c-MYC expression does not prevent P/CQ-mediated cell cycle arrest. (A) Panc 10.05, PDX220, and MIA-PaCa2 cells were imaged to assess confluence over time using the IncuCyte Live-Cell Analysis System after treatment with palbociclib (P), CQ, or the combination of both agents (P/CQ). Data represent mean values, and error bars reflect SD of one experiment ( n = 3). One-way ANOVA was conducted to determine statistical difference between the single agent controls (CDK4/6 inhibitor or CQ) and the combination group. *, P < 0.05; **, P < 0.01; ***, P < 0.001. (B) MIA-PaCa2 cells were treated with ribociclib (Ribo) or abemaciclib (Abema) alone or in combination with CQ and confluence was measured over time. A one-way ANOVA was utilized to determine statistical significance between the single agent controls and the combination group. The Ribo/CQ experiment was conducted twice ( n = 2) and the Abema/CQ experiment was conducted once ( n = 3). *, P < 0.05; **, P < 0.01; ***, P < 0.001. (C) Representative images of MIA-PaCa2 cells shown by phase contrast (top) and the corresponding cell mask utilized to estimate well confluence (bottom) for each treatment at 48 h. Bars are 300 μm. (D) Average percentage of cells in S phase for Tet-On c-MYC–expressing Panc 10.05, HPAF-II, and PDX220 cells that were treated for 7 d with DMSO (Ctrl); 2 μg/ml Dox; 5–10 μM CQ and 2.5 μM palbociclib (P/CQ); or D/P/CQ. Control and Dox-treated sample percentages are continued from ; experiments were performed at the same time as P/CQ- and D/P/CQ-treated samples ( n = 3 from two independent experiments, mean ± SEM). *, P < 0.05; **, P < 0.01; ns, not significant by Student’s two-sided t test. (E) Immunoblot analysis of lysates generated from Tet-On c-MYC–expressing cells that were treated for 7 d with DMSO (Ctrl), 2 μg/ml Dox (D), 5–10 μM CQ, 2.5 μM palbociclib (P), or the indicated combinations (representative blots from two independent experiments). Lysates were blotted for c-MYC, pRB (phospho-S807/811), RB, CCNB1, CCNA2, and ACTB. Densitometry quantitation of c-MYC and pRB protein levels normalized to actin are shown. Source data are available for this figure: .

    Article Snippet: Antibodies used are as follows (used at 1:1,000 unless noted otherwise): 1:2,000 phospho-T202/Y204 p44/42 MAPK (ERK1/2; CST:4370), total ERK1/2 (CST:4696); p-RB 807/811 1:1,000 (CST 8516S); RB (CST:9309S); c-MYC (CST:5605S); E2F1 (CST:3742S); Cyclin B1 (CST:12231S); Cyclin A2 (CST:91500S); Cyclin D2 (CST:3741S); Cyclin E1 (CST:20808S); CDK2 (CST:2546S); CDK4 (CST:12790S); CDK6 (CST:3136S); p27 KIP1 , (CST:3686S); p21 WAF1/CIP1 (CST:2947S); 1:1,000 Vinculin (CST:13901); 1:2,000 GAPDH (CST:97166S); 1:5,000 β-actin (CST:4970S).

    Techniques: Expressing, Western Blot, Generated, Quantitation Assay

    The qPCR primers of different genes

    Journal: American Journal of Cancer Research

    Article Title: DOT1L promotes cell proliferation and invasion by epigenetically regulating STAT5B in renal cell carcinoma

    doi:

    Figure Lengend Snippet: The qPCR primers of different genes

    Article Snippet: Antibodies against cyclin A2 (CCNA2; #91500), Bcl2-associated X protein (Bax; #5023), and phospho-STAT5 (Tyr694) (#4322) were purchased from Cell Signaling Technology (Massachusetts, USA).

    Techniques:

    Inhibition or knockdown of DOT1L induces S-phase arrest in 786-O cells. A. The cell cycle was analyzed by flow cytometry in 786-O cells treated with SGC0946 (left). Cell cycle data are presented as the means ± SDs (right). B. The cell cycle was analyzed by flow cytometry in 786-O cells after DOT1L knockdown (left). Cell cycle data are presented as the means ± SDs (right). C. The mRNA levels of cell cycle-related proteins, including cyclin A1, A2, B1, D1, D2, and E2; CDK2, CDK4, and CDK6; and P21, p53, and RB, were measured by qPCR in 786-O cells after DOT1L knockdown. D. The protein levels of selected cell cycle-related proteins (P21, cyclin A2, and CDK6) were measured by using western blotting in DOT1L knockdown 786-O cells. *P<0.05; **P<0.01; ***P<0.001 compared with the 0 µM group or control group.

    Journal: American Journal of Cancer Research

    Article Title: DOT1L promotes cell proliferation and invasion by epigenetically regulating STAT5B in renal cell carcinoma

    doi:

    Figure Lengend Snippet: Inhibition or knockdown of DOT1L induces S-phase arrest in 786-O cells. A. The cell cycle was analyzed by flow cytometry in 786-O cells treated with SGC0946 (left). Cell cycle data are presented as the means ± SDs (right). B. The cell cycle was analyzed by flow cytometry in 786-O cells after DOT1L knockdown (left). Cell cycle data are presented as the means ± SDs (right). C. The mRNA levels of cell cycle-related proteins, including cyclin A1, A2, B1, D1, D2, and E2; CDK2, CDK4, and CDK6; and P21, p53, and RB, were measured by qPCR in 786-O cells after DOT1L knockdown. D. The protein levels of selected cell cycle-related proteins (P21, cyclin A2, and CDK6) were measured by using western blotting in DOT1L knockdown 786-O cells. *P<0.05; **P<0.01; ***P<0.001 compared with the 0 µM group or control group.

    Article Snippet: Antibodies against cyclin A2 (CCNA2; #91500), Bcl2-associated X protein (Bax; #5023), and phospho-STAT5 (Tyr694) (#4322) were purchased from Cell Signaling Technology (Massachusetts, USA).

    Techniques: Inhibition, Flow Cytometry, Western Blot

    Overexpression of STAT5B partially rescues DOT1L silencing-induced cell proliferation inhibition and cell cycle arrest. A. Assessment of the DNA transfection efficiency after incubation with the pEGFP-N1/STAT5B vector (oeSTAT5B) or the pEGFP-N1 (sham) vector in 786-O cells with sh2-mediated DOT1L knockdown at the 24 h time point. B. DOT1L and STAT5B mRNA expression was measured by using qPCR in 786-O cells after DOT1L silencing and STAT5B overexpression. C. DOT1L, STAT5B and p-STAT5 protein levels were measured by Western blotting in 786-O cells after DOT1L knockdown and STAT5B overexpression. D. The cell growth curve was constructed with CCK8 assay data in 786-O cells after DOT1L knockdown and STAT5B overexpression. Both the sh2 and sh2+sham groups were used as the vector controls for STAT5B overexpression at the 0/24/48/72 h time points. E. The cell cycle was analyzed by flow cytometry in 786-O cells with DOT1L knockdown and STAT5B restoration. F. Cyclin A2, P21, and CDK6 protein expression was measured by Western blotting in 786-O cells after DOT1L knockdown and STAT5B overexpression.

    Journal: American Journal of Cancer Research

    Article Title: DOT1L promotes cell proliferation and invasion by epigenetically regulating STAT5B in renal cell carcinoma

    doi:

    Figure Lengend Snippet: Overexpression of STAT5B partially rescues DOT1L silencing-induced cell proliferation inhibition and cell cycle arrest. A. Assessment of the DNA transfection efficiency after incubation with the pEGFP-N1/STAT5B vector (oeSTAT5B) or the pEGFP-N1 (sham) vector in 786-O cells with sh2-mediated DOT1L knockdown at the 24 h time point. B. DOT1L and STAT5B mRNA expression was measured by using qPCR in 786-O cells after DOT1L silencing and STAT5B overexpression. C. DOT1L, STAT5B and p-STAT5 protein levels were measured by Western blotting in 786-O cells after DOT1L knockdown and STAT5B overexpression. D. The cell growth curve was constructed with CCK8 assay data in 786-O cells after DOT1L knockdown and STAT5B overexpression. Both the sh2 and sh2+sham groups were used as the vector controls for STAT5B overexpression at the 0/24/48/72 h time points. E. The cell cycle was analyzed by flow cytometry in 786-O cells with DOT1L knockdown and STAT5B restoration. F. Cyclin A2, P21, and CDK6 protein expression was measured by Western blotting in 786-O cells after DOT1L knockdown and STAT5B overexpression.

    Article Snippet: Antibodies against cyclin A2 (CCNA2; #91500), Bcl2-associated X protein (Bax; #5023), and phospho-STAT5 (Tyr694) (#4322) were purchased from Cell Signaling Technology (Massachusetts, USA).

    Techniques: Over Expression, Inhibition, Transfection, Incubation, Plasmid Preparation, Expressing, Western Blot, Construct, CCK-8 Assay, Flow Cytometry