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plko 1 shtgfbr3 puro  (Addgene inc)


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    Addgene inc plko 1 shtgfbr3 puro
    Fig. 3. TGFBR3 activates BCL2-family-dependent apoptotic signaling. (A) Relative basal mRNA levels of BCL2L1 in MCF10A-5E cells that were transduced with pBABE-neo or TGFBR3-HA. mRNA levels were analyzed by qRT-PCR. BCL2L1 transcripts were normalized so that the geometric mean of relative abundance of BCL2L1 in control cells equals one. (B–E) Relative mRNA levels of BH3-only BCL-2 family members [BBC3 (B), BAD (C), BCL1L11 (D), and BID (E)]. MCF10A-5E cells expressing shGFP, <t>shTGFBR3#1</t> or shTGFBR3#2 were placed in suspension for the indicated times, and population-level mRNA measurements were performed by qRT-PCR. Transcripts were normalized so that the geometric mean of relative abundance of the indicated transcript in shGFP-expressing cells at time 0 equals one. (F) Relative basal mRNA levels of BBC3 and BAD in MCF10A-5E cells that were transduced with pBABE-neo or TGFBR3-HA. mRNA levels were analyzed by qRT-PCR. Transcripts were normalized so that the geometric mean of relative abundance of the indicated transcript in control cells equals one. For A–F, data are shown as the mean±s.e.m. of four independent biological samples. P-values were calculated by Welch’s two-sided t-test; n.s., not significant.
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    1) Product Images from "TGFBR3 supports anoikis through suppressing ATF4 signaling."

    Article Title: TGFBR3 supports anoikis through suppressing ATF4 signaling.

    Journal: Journal of cell science

    doi: 10.1242/jcs.258396

    Fig. 3. TGFBR3 activates BCL2-family-dependent apoptotic signaling. (A) Relative basal mRNA levels of BCL2L1 in MCF10A-5E cells that were transduced with pBABE-neo or TGFBR3-HA. mRNA levels were analyzed by qRT-PCR. BCL2L1 transcripts were normalized so that the geometric mean of relative abundance of BCL2L1 in control cells equals one. (B–E) Relative mRNA levels of BH3-only BCL-2 family members [BBC3 (B), BAD (C), BCL1L11 (D), and BID (E)]. MCF10A-5E cells expressing shGFP, shTGFBR3#1 or shTGFBR3#2 were placed in suspension for the indicated times, and population-level mRNA measurements were performed by qRT-PCR. Transcripts were normalized so that the geometric mean of relative abundance of the indicated transcript in shGFP-expressing cells at time 0 equals one. (F) Relative basal mRNA levels of BBC3 and BAD in MCF10A-5E cells that were transduced with pBABE-neo or TGFBR3-HA. mRNA levels were analyzed by qRT-PCR. Transcripts were normalized so that the geometric mean of relative abundance of the indicated transcript in control cells equals one. For A–F, data are shown as the mean±s.e.m. of four independent biological samples. P-values were calculated by Welch’s two-sided t-test; n.s., not significant.
    Figure Legend Snippet: Fig. 3. TGFBR3 activates BCL2-family-dependent apoptotic signaling. (A) Relative basal mRNA levels of BCL2L1 in MCF10A-5E cells that were transduced with pBABE-neo or TGFBR3-HA. mRNA levels were analyzed by qRT-PCR. BCL2L1 transcripts were normalized so that the geometric mean of relative abundance of BCL2L1 in control cells equals one. (B–E) Relative mRNA levels of BH3-only BCL-2 family members [BBC3 (B), BAD (C), BCL1L11 (D), and BID (E)]. MCF10A-5E cells expressing shGFP, shTGFBR3#1 or shTGFBR3#2 were placed in suspension for the indicated times, and population-level mRNA measurements were performed by qRT-PCR. Transcripts were normalized so that the geometric mean of relative abundance of the indicated transcript in shGFP-expressing cells at time 0 equals one. (F) Relative basal mRNA levels of BBC3 and BAD in MCF10A-5E cells that were transduced with pBABE-neo or TGFBR3-HA. mRNA levels were analyzed by qRT-PCR. Transcripts were normalized so that the geometric mean of relative abundance of the indicated transcript in control cells equals one. For A–F, data are shown as the mean±s.e.m. of four independent biological samples. P-values were calculated by Welch’s two-sided t-test; n.s., not significant.

    Techniques Used: Transduction, Quantitative RT-PCR, Control, Expressing, Suspension



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    Fig. 3. TGFBR3 activates BCL2-family-dependent apoptotic signaling. (A) Relative basal mRNA levels of BCL2L1 in MCF10A-5E cells that were transduced with pBABE-neo or TGFBR3-HA. mRNA levels were analyzed by qRT-PCR. BCL2L1 transcripts were normalized so that the geometric mean of relative abundance of BCL2L1 in control cells equals one. (B–E) Relative mRNA levels of BH3-only BCL-2 family members [BBC3 (B), BAD (C), BCL1L11 (D), and BID (E)]. MCF10A-5E cells expressing shGFP, <t>shTGFBR3#1</t> or shTGFBR3#2 were placed in suspension for the indicated times, and population-level mRNA measurements were performed by qRT-PCR. Transcripts were normalized so that the geometric mean of relative abundance of the indicated transcript in shGFP-expressing cells at time 0 equals one. (F) Relative basal mRNA levels of BBC3 and BAD in MCF10A-5E cells that were transduced with pBABE-neo or TGFBR3-HA. mRNA levels were analyzed by qRT-PCR. Transcripts were normalized so that the geometric mean of relative abundance of the indicated transcript in control cells equals one. For A–F, data are shown as the mean±s.e.m. of four independent biological samples. P-values were calculated by Welch’s two-sided t-test; n.s., not significant.
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    Fig. 3. TGFBR3 activates BCL2-family-dependent apoptotic signaling. (A) Relative basal mRNA levels of BCL2L1 in MCF10A-5E cells that were transduced with pBABE-neo or TGFBR3-HA. mRNA levels were analyzed by qRT-PCR. BCL2L1 transcripts were normalized so that the geometric mean of relative abundance of BCL2L1 in control cells equals one. (B–E) Relative mRNA levels of BH3-only BCL-2 family members [BBC3 (B), BAD (C), BCL1L11 (D), and BID (E)]. MCF10A-5E cells expressing shGFP, <t>shTGFBR3#1</t> or shTGFBR3#2 were placed in suspension for the indicated times, and population-level mRNA measurements were performed by qRT-PCR. Transcripts were normalized so that the geometric mean of relative abundance of the indicated transcript in shGFP-expressing cells at time 0 equals one. (F) Relative basal mRNA levels of BBC3 and BAD in MCF10A-5E cells that were transduced with pBABE-neo or TGFBR3-HA. mRNA levels were analyzed by qRT-PCR. Transcripts were normalized so that the geometric mean of relative abundance of the indicated transcript in control cells equals one. For A–F, data are shown as the mean±s.e.m. of four independent biological samples. P-values were calculated by Welch’s two-sided t-test; n.s., not significant.
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    Fig. 3. TGFBR3 activates BCL2-family-dependent apoptotic signaling. (A) Relative basal mRNA levels of BCL2L1 in MCF10A-5E cells that were transduced with pBABE-neo or TGFBR3-HA. mRNA levels were analyzed by qRT-PCR. BCL2L1 transcripts were normalized so that the geometric mean of relative abundance of BCL2L1 in control cells equals one. (B–E) Relative mRNA levels of BH3-only BCL-2 family members [BBC3 (B), BAD (C), BCL1L11 (D), and BID (E)]. MCF10A-5E cells expressing shGFP, <t>shTGFBR3#1</t> or shTGFBR3#2 were placed in suspension for the indicated times, and population-level mRNA measurements were performed by qRT-PCR. Transcripts were normalized so that the geometric mean of relative abundance of the indicated transcript in shGFP-expressing cells at time 0 equals one. (F) Relative basal mRNA levels of BBC3 and BAD in MCF10A-5E cells that were transduced with pBABE-neo or TGFBR3-HA. mRNA levels were analyzed by qRT-PCR. Transcripts were normalized so that the geometric mean of relative abundance of the indicated transcript in control cells equals one. For A–F, data are shown as the mean±s.e.m. of four independent biological samples. P-values were calculated by Welch’s two-sided t-test; n.s., not significant.
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    Fig. 3. TGFBR3 activates BCL2-family-dependent apoptotic signaling. (A) Relative basal mRNA levels of BCL2L1 in MCF10A-5E cells that were transduced with pBABE-neo or TGFBR3-HA. mRNA levels were analyzed by qRT-PCR. BCL2L1 transcripts were normalized so that the geometric mean of relative abundance of BCL2L1 in control cells equals one. (B–E) Relative mRNA levels of BH3-only BCL-2 family members [BBC3 (B), BAD (C), BCL1L11 (D), and BID (E)]. MCF10A-5E cells expressing shGFP, shTGFBR3#1 or shTGFBR3#2 were placed in suspension for the indicated times, and population-level mRNA measurements were performed by qRT-PCR. Transcripts were normalized so that the geometric mean of relative abundance of the indicated transcript in shGFP-expressing cells at time 0 equals one. (F) Relative basal mRNA levels of BBC3 and BAD in MCF10A-5E cells that were transduced with pBABE-neo or TGFBR3-HA. mRNA levels were analyzed by qRT-PCR. Transcripts were normalized so that the geometric mean of relative abundance of the indicated transcript in control cells equals one. For A–F, data are shown as the mean±s.e.m. of four independent biological samples. P-values were calculated by Welch’s two-sided t-test; n.s., not significant.

    Journal: Journal of cell science

    Article Title: TGFBR3 supports anoikis through suppressing ATF4 signaling.

    doi: 10.1242/jcs.258396

    Figure Lengend Snippet: Fig. 3. TGFBR3 activates BCL2-family-dependent apoptotic signaling. (A) Relative basal mRNA levels of BCL2L1 in MCF10A-5E cells that were transduced with pBABE-neo or TGFBR3-HA. mRNA levels were analyzed by qRT-PCR. BCL2L1 transcripts were normalized so that the geometric mean of relative abundance of BCL2L1 in control cells equals one. (B–E) Relative mRNA levels of BH3-only BCL-2 family members [BBC3 (B), BAD (C), BCL1L11 (D), and BID (E)]. MCF10A-5E cells expressing shGFP, shTGFBR3#1 or shTGFBR3#2 were placed in suspension for the indicated times, and population-level mRNA measurements were performed by qRT-PCR. Transcripts were normalized so that the geometric mean of relative abundance of the indicated transcript in shGFP-expressing cells at time 0 equals one. (F) Relative basal mRNA levels of BBC3 and BAD in MCF10A-5E cells that were transduced with pBABE-neo or TGFBR3-HA. mRNA levels were analyzed by qRT-PCR. Transcripts were normalized so that the geometric mean of relative abundance of the indicated transcript in control cells equals one. For A–F, data are shown as the mean±s.e.m. of four independent biological samples. P-values were calculated by Welch’s two-sided t-test; n.s., not significant.

    Article Snippet: pLKO.1 shTGFBR3 puro (Addgene #58696, shTGFBR3#1) and pBabe neo TGFBR3-HA (Addgene #83095) were previously described (Wang et al., 2014; Bajikar et al., 2017). pLKO.1 puro (Addgene #8453), Tet-pLKO-neo (Addgene #21916), pLKO.1 GFP shRNA (Addgene #30323) (Sancak et al., 2008), pBABE-neo (Addgene #1767) (Morgenstern and Land, 1990), pDONR223 LacZ (Addgene #25893), pDONR223_ATF4_WT (Addgene #82190), and pLX304 (Addgene #25890) (Yang et al., 2011) were obtained commercially.

    Techniques: Transduction, Quantitative RT-PCR, Control, Expressing, Suspension

    KEY RESOURCES TABLE

    Journal: Developmental cell

    Article Title: Tumor Suppressor Inactivation of GDF11 Occurs by Precursor Sequestration in Triple-Negative Breast Cancer

    doi: 10.1016/j.devcel.2017.10.027

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: Cross) N/A Experimental Models: Organisms/Strains Mouse: SCID Beige (CB17.Cg-Prkdc scid Lyst bg-J /Crl) Charles River Strain #250, RRID: IMSR_CRL:250 Mouse: BALB/c (BALB/cAnNCrl) Charles River Strain #028, RRID: IMSR_CRL:28 Oligonucleotides Primers for quantitative PCR, see Table S6 This paper N/A Recombinant DNA TET-pLKO.1 puro shLuc This paper Addgene #83092 TET-pLKO.1 puro shGFP This paper Addgene #83085 TET-pLKO.1 puro shLacZ This paper Addgene #98632 TET-pLKO.1 puro shGDF11 #1 This paper Addgene #83083 TET-pLKO.1 puro shGDF11 #2 This paper Addgene #83084 TET-pLKO.1 puro shSMAD4 #1 This paper Addgene #83089 TET-pLKO.1 puro shSMAD4 #2 This paper Addgene #83091 TET-pLKO.1 puro shID2 #1 This paper Addgene #83086 TET-pLKO.1 puro shID2 #2 This paper Addgene #83087 TET-pLKO.1 puro shId2 #1 This paper Addgene #83088 TET-pLKO.1 puro shId2 #2 This paper Addgene #83090 pLX304 LacZ-2′V5 blast This paper Addgene #83099 pLX304 GDF11-V5 blast This paper Addgene #83098 pLX304 PCSK5-V5 blast This paper Addgene #83100 pLX304 PCSK5 (T288P)-V5 blast This paper Addgene #83101 pLX304 PCSK5 (A270T)-V5 blast This paper Addgene #83102 pLX304 PCSK5 (H516P)-V5 blast This paper Addgene #83103 pLX304 PCSK5 (R511C)-V5 blast This paper Addgene #83104 pLX304 Luciferase-V5 blast This paper Addgene #98580 pLX302 ID2-V5 puro This paper Addgene #83096 pLX302 GDF11-V5 puro This paper Addgene #83097 pLX302 PCSK5-V5 puro This paper Addgene #98389 pLX302 PCSK5 (T288P)-V5 puro This paper Addgene #98709 pLX302 LacZ-2′V5 blast This paper Addgene #98579 pEN_TT_miRc2 3xFLAG This paper Addgene #83274 pEN_TT_miRc2 3xFLAG LacZ This paper Addgene #83093 pEN_TT_miRc2 3xFLAG Luciferase This paper Addgene #98391 pEN_TT_miRc2 3XFLAG SMAD4 This paper Addgene #83094 pEN_TT_miRc2 3XFLAG ID2 This paper Addgene #98390 pSLIK_TT 3xFLAG LacZ neo This paper Addgene #83105 pSLIK_TT 3xFLAG LacZ hygro This paper Addgene #98395 pSLIK_TT 3xFLAG Luciferase neo This paper Addgene #98392 pSLIK_TT 3xFLAG SMAD4 neo This paper Addgene #83273 pSLIK_TT 3xFLAG ID2 neo This paper Addgene #98393 pSLIK_TT 3xFLAG ID2 hygro This paper Addgene #98394 pBabe TGFBR3-HA neo This paper Addgene #83095 pBabe Venus neo This paper Addgene #98396 pBabe mRFP1 neo This paper Addgene #98397 pLKO.1 shTGFBR3 puro Wang et al., 2014 .

    Techniques: In Situ, Recombinant, Enzyme-linked Immunosorbent Assay, Mutagenesis, Amplification, Expressing, Real-time Polymerase Chain Reaction, Luciferase, Software

    KEY RESOURCES TABLE

    Journal: Developmental cell

    Article Title: Tumor Suppressor Inactivation of GDF11 Occurs by Precursor Sequestration in Triple-Negative Breast Cancer

    doi: 10.1016/j.devcel.2017.10.027

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: Plasmids pLKO.1 shTGFBR3 puro (Addgene #58696), pBabe TGFBR3-HA neo, and pLX302 luciferase-V5 puro (Addgene #47553) were previously described ( Kang et al., 2013 ; Wang et al., 2014 ). pBabe RasV12 puro (Addgene #1768), pLenti PGK V5-luciferase (w528-1) blast (Addgene #19166) ( Campeau et al., 2009 ), pDONR223 LacZ (Addgene #25893) ( Yang et al., 2011 ), pDONR223 luciferase (Addgene #25894) ( Yang et al., 2011 ), pENTR223.1 (pro)GDF11 (DNASU # HsCD00351455, Arizona State), and pCMV-SPORT6 Id2 (#2655173, Dharmacon) were obtained commercially. pDONR223 plasmids for SMAD4, ID2 and PC5A (PCSK5) were pulled from the human ORFeome V5.1 ( Yang et al., 2011 ). shRNA targeting sequences from the RNAi Consortium ( Moffat et al., 2006 ) were cloned into tet-pLKO.1 puro ( Wiederschain et al., 2009 ) (Addgene #21915) exactly as previously described for pLKO.1 puro ( Bajikar et al., 2014 ): shGFP (sequence from Addgene #30323), shLuc (TRCN0000072243), shLacZ (TRCN0000072238), shGDF11 #1 (TRCN0000438352), shGDF11 #2 (TRCN0000442158), shSMAD4 #1 (TRCN0000025900), shSMAD4 #2 (TRCN0000288653), shID2 #1 (TRCN0000232080), shID2 #2 (TRCN0000232079), shId2 #1 (TRCN0000229535), and shId2 #2 (TRCN0000218289).

    Techniques: Virus, In Situ, Recombinant, Enzyme-linked Immunosorbent Assay, Mutagenesis, RNA Amplification, Expressing, Real-time Polymerase Chain Reaction, Luciferase, Software

    KEY RESOURCES TABLE

    Journal: Developmental cell

    Article Title: Tumor Suppressor Inactivation of GDF11 Occurs by Precursor Sequestration in Triple-Negative Breast Cancer

    doi: 10.1016/j.devcel.2017.10.027

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: Cross) N/A Experimental Models: Organisms/Strains Mouse: SCID Beige (CB17.Cg-Prkdc scid Lyst bg-J /Crl) Charles River Strain #250, RRID: IMSR_CRL:250 Mouse: BALB/c (BALB/cAnNCrl) Charles River Strain #028, RRID: IMSR_CRL:28 Oligonucleotides Primers for quantitative PCR, see Table S6 This paper N/A Recombinant DNA TET-pLKO.1 puro shLuc This paper Addgene #83092 TET-pLKO.1 puro shGFP This paper Addgene #83085 TET-pLKO.1 puro shLacZ This paper Addgene #98632 TET-pLKO.1 puro shGDF11 #1 This paper Addgene #83083 TET-pLKO.1 puro shGDF11 #2 This paper Addgene #83084 TET-pLKO.1 puro shSMAD4 #1 This paper Addgene #83089 TET-pLKO.1 puro shSMAD4 #2 This paper Addgene #83091 TET-pLKO.1 puro shID2 #1 This paper Addgene #83086 TET-pLKO.1 puro shID2 #2 This paper Addgene #83087 TET-pLKO.1 puro shId2 #1 This paper Addgene #83088 TET-pLKO.1 puro shId2 #2 This paper Addgene #83090 pLX304 LacZ-2′V5 blast This paper Addgene #83099 pLX304 GDF11-V5 blast This paper Addgene #83098 pLX304 PCSK5-V5 blast This paper Addgene #83100 pLX304 PCSK5 (T288P)-V5 blast This paper Addgene #83101 pLX304 PCSK5 (A270T)-V5 blast This paper Addgene #83102 pLX304 PCSK5 (H516P)-V5 blast This paper Addgene #83103 pLX304 PCSK5 (R511C)-V5 blast This paper Addgene #83104 pLX304 Luciferase-V5 blast This paper Addgene #98580 pLX302 ID2-V5 puro This paper Addgene #83096 pLX302 GDF11-V5 puro This paper Addgene #83097 pLX302 PCSK5-V5 puro This paper Addgene #98389 pLX302 PCSK5 (T288P)-V5 puro This paper Addgene #98709 pLX302 LacZ-2′V5 blast This paper Addgene #98579 pEN_TT_miRc2 3xFLAG This paper Addgene #83274 pEN_TT_miRc2 3xFLAG LacZ This paper Addgene #83093 pEN_TT_miRc2 3xFLAG Luciferase This paper Addgene #98391 pEN_TT_miRc2 3XFLAG SMAD4 This paper Addgene #83094 pEN_TT_miRc2 3XFLAG ID2 This paper Addgene #98390 pSLIK_TT 3xFLAG LacZ neo This paper Addgene #83105 pSLIK_TT 3xFLAG LacZ hygro This paper Addgene #98395 pSLIK_TT 3xFLAG Luciferase neo This paper Addgene #98392 pSLIK_TT 3xFLAG SMAD4 neo This paper Addgene #83273 pSLIK_TT 3xFLAG ID2 neo This paper Addgene #98393 pSLIK_TT 3xFLAG ID2 hygro This paper Addgene #98394 pBabe TGFBR3-HA neo This paper Addgene #83095 pBabe Venus neo This paper Addgene #98396 pBabe mRFP1 neo This paper Addgene #98397 pLKO.1 shTGFBR3 puro Wang et al., 2014 .

    Techniques: In Situ, Recombinant, Enzyme-linked Immunosorbent Assay, Mutagenesis, Amplification, Expressing, Real-time Polymerase Chain Reaction, Luciferase, Software