Journal: The Journal of Neuroscience

Article Title: The Ubiquitin E3 Ligase TRAF6 Exacerbates Ischemic Stroke by Ubiquitinating and Activating Rac1

doi: 10.1523/JNEUROSCI.1751-17.2017

Figure Lengend Snippet: Antibodies used for immunoblot analyses

Article Snippet: The protein bands were detected using a Bio-Rad imaging system and specific protein expression levels were normalized to GAPDH protein. table ft1 table-wrap mode="anchored" t5 Table 3. caption a7 Antibody Manufacturer Catalog # Sources of species RRID TRAF6 Abcam Ab40675 Rabbit AB_778573 Bax CST 2772 Rabbit AB_329921 Bcl-2(50E 3 ) CST 2870 Rabbit AB_2290370 Cleaved Caspase3(Asp175) CST 9661 Rabbit AB_2341188 Caspase-3 CST 9662 Rabbit AB_331439 p-IkBa(ser32/36) CST 9246L Mouse AB_2267145 IkBa(L35A5) CST 4814 Mouse AB_390781 P-Ikkβ(Y199) Abcam Ab59195 Rabbit AB_943810 IKKβ CST 8943S Rabbit AB_11024092 P-NF-kB p65(Ser536) CST 3033 Rabbit AB_331284 p65 CST 4764 Rabbit AB_823578 P47-phox(P366) Bioworld BS3261 Rabbit AB_1662897 NOXA2/P67 phox Abcam {"type":"entrez-nucleotide","attrs":{"text":"Ab109366","term_id":"37572651","term_text":"AB109366"}} Ab109366 Rabbit AB_10860640 SD1(SOD1) Abcam Ab13498 Rabbit AB_300402 Heme Oxygenase 1(HO-1) Abcam Ab13243 Rabbit AB_299790 p-Rac1/cdc42(Ser71) CST 2461S Rabbit AB_2300703 Rac1/2/3 CST 2467 Rabbit AB_2176150 HA Sigma-Aldrich H6908 Rabbit AB_260070 Flag Sigma-Aldrich F3165 Mouse AB_259529 Myc MBL M192-3 Mouse AB_11160947 Ub CST 3933S Rabbit AB_2180538 His CST 12698 Rabbit Gst CST 2624S Mouse AB_10692101 GAPDH(14C10) CST 2118 Rabbit AB_561053 Open in a separate window Antibodies used for immunoblot analyses Cell culture and in vitro manipulation.

Techniques: Western Blot

Journal: Nature Communications

Article Title: The nuclear receptor ERR cooperates with the cardiogenic factor GATA4 to orchestrate cardiomyocyte maturation

doi: 10.1038/s41467-022-29733-3

Figure Lengend Snippet: a Real-time quantitative polymerase chain reaction (RT-qPCR)-determined levels of the designated mRNAs during human induced pluripotent stem cell-derived cardiomyocyte (hiPSC-CM) differentiation. Levels of indicated genes are shown normalized to RPLP0 levels at each timepoint. Day 0 and 2, n = 9; Day 4, 6, 8, and 15, n = 8. * p < 0.05, *** p < 0.001, **** p < 0.0001 vs Day 0; one-way ANOVA followed by Dunnett’s multiple comparison test. b Representative immunoblots of estrogen-related receptor (ERR) α and γ protein levels during hiPSC-CM differentiation. α-Tubulin was used as a loading control. c Heatmap representing log 2 fold change [ERRα/γ knockout (KO)/wild-type control (WT)] of mRNA levels of genes involved in cardiac structural component, Ca 2+ handling function, and ion transport in two different KO cell lines (KO1, KO6). All changes were significant (Benjamini-Hochberg false discovery rate <0.05) except for SCN5A in KO1 ( p = 0.057). d Representative immunoblots of indicated protein levels in WT Control and ERRα/γ KO hiPSC-CMs. Total protein staining was shown as a loading control. e The mRNA levels of indicated genes determined by RT-qPCR in hiPSC-CMs following the infection of adenovirus expressing GFP or ERRγ (Ad-GFP or Ad-ERRγ). ** p < 0.01, *** p < 0.001, **** p < 0.0001, Ad-ERRγ vs Ad-GFP, two-tailed student’s t-test. PDK4 (Ad-GFP, n = 7 and Ad-ERRγ, n = 8); COX6A2 (Ad-GFP, n = 14 and Ad-ERRγ, n = 15); MDH1 and CKMT2 , (Ad-GFP, n = 7 and Ad-ERRγ, n = 8); TNNI3 (Ad-GFP, n = 14 and Ad-ERRγ, n = 15); TNNI1 (Ad-GFP, n = 10 and Ad-ERRγ, n = 11). f The mRNA levels of indicated hiPSC-CM maturation markers determined by RT-qPCR in WT and ERRα/γ KO hiPSC-CMs derived from KO1 and KO6 with ( n = 15) or without ( n = 9) maturation cocktail treatment for 7 days. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, WT vs ERRα/γ KO hiPSC-CMs, two-way ANOVA followed by Tukey’s multiple comparisons test. All graphs in a , e , and f represent the means ± SEM. n denotes independent biological replicates. g Representative immunoblot images to show ACSL1 and TNNI3 protein expression levels in WT and ERRα/γ KO hiPSC-CMs with or without maturation cocktail. Total protein staining was used as a loading control.

Article Snippet: The following antibodies were used: ERRα; SMYD1; VDAC; ACSL3; TNNI3 (Abcam catalog and clone #s: ab76228; EPR46Y, ab181372; EPR13574(B)-30, ab15895, ab151959, and ab47003 respectively); His tag; α-tubulin; ACSL1 (Cell Signaling Technology catalog and clone #s: 12698S; D3I1O, 3873; DM1A, and 9189; D2H5 respectively); HA tag and MYL2 (Proteintech catalog #s 51064-2-AP and 10906-1-AP); α-Actinin, FLAG M2, TNNI1, ACTB, and PGC-1α (Sigma-Aldrich catalog and clone #s: A7732-100UL; EA-53, F1804-50UG; M2, AV42117-100UL, A5316; AC-74, and ST1202; 4C1.3); GATA4 and MYBPC3 (SANTA CRUZ BIOTECHNOLOGY catalog and clone #s, sc-25310; G-4, and sc-137180; E-7).

Techniques: Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Derivative Assay, Comparison, Western Blot, Control, Knock-Out, Staining, Infection, Expressing, Two Tailed Test