Journal: BMC Immunology

Article Title: Establishment and validation of a recurrent prediction model for glioma: extrinsic apoptotic molecules FADD and CASP8 are closely associated with glioma recurrence

doi: 10.1186/s12865-025-00746-z

Figure Lengend Snippet: The association between recurrent score and classical apoptotic genes. (A) The relationship between the 6 genes and recurrent score in CGGA and TCGA database . (B) PPI network of CASP3, CASP9, FADD, CASP7, CASP8, BCL2，and the 9-gene signature from the STRING. (C-D) The expression levels of the 6 apoptotic genes in low- and high-risk levels . (E-J) Correlation between recurrent score and expression levels of apoptotic genes. *P<0.05; ***P<0.001; ns, not significant

Article Snippet: Characterizing the differential expression patterns of CASP8 and FADD in gliomas and normal tissues will play a crucial role in the further development of targeted therapeutic strategies for gliomas Fig. 9 RNA and protein levels of CASP8 and FADD in normal tissues and tumors. (A-B) RNA expression of CASP8 and FADD in normal tissues from the NCBI database (https://www.ncbi.nlm.nih.gov/). (C-D) RNA expression of CASP8 and FADD in normal tissues from the Human Protein Atlas database (https://www.proteinatlas.org/). (E) Protein levels of CASP8 and FADD in normal brain tissues from The Human Protein Atlas database. (F) Protein levels of CASP8 and FADD in normal tissues from The Human Protein Atlas database. (G) Protein levels of CASP8 and FADD in tumors from The Human Protein Atlas database

Techniques: Expressing

Journal: BMC Immunology

Article Title: Establishment and validation of a recurrent prediction model for glioma: extrinsic apoptotic molecules FADD and CASP8 are closely associated with glioma recurrence

doi: 10.1186/s12865-025-00746-z

Figure Lengend Snippet: RNA and protein levels of CASP8 and FADD in normal tissues and tumors. (A-B) RNA expression of CASP8 and FADD in normal tissues from the NCBI database (https://www.ncbi.nlm.nih.gov/). (C-D) RNA expression of CASP8 and FADD in normal tissues from the Human Protein Atlas database (https://www.proteinatlas.org/). (E) Protein levels of CASP8 and FADD in normal brain tissues from The Human Protein Atlas database. (F) Protein levels of CASP8 and FADD in normal tissues from The Human Protein Atlas database. (G) Protein levels of CASP8 and FADD in tumors from The Human Protein Atlas database

Article Snippet: Characterizing the differential expression patterns of CASP8 and FADD in gliomas and normal tissues will play a crucial role in the further development of targeted therapeutic strategies for gliomas Fig. 9 RNA and protein levels of CASP8 and FADD in normal tissues and tumors. (A-B) RNA expression of CASP8 and FADD in normal tissues from the NCBI database (https://www.ncbi.nlm.nih.gov/). (C-D) RNA expression of CASP8 and FADD in normal tissues from the Human Protein Atlas database (https://www.proteinatlas.org/). (E) Protein levels of CASP8 and FADD in normal brain tissues from The Human Protein Atlas database. (F) Protein levels of CASP8 and FADD in normal tissues from The Human Protein Atlas database. (G) Protein levels of CASP8 and FADD in tumors from The Human Protein Atlas database

Techniques: RNA Expression

Journal: Pharmacognosy magazine

Article Title: Poncirus trifoliata Rafin. induces the apoptosis of triple-negative breast cancer cells via activation of the c-Jun NH(2)-terminal kinase and extracellular signal-regulated kinase pathways.

doi: 10.4103/0973-1296.166056

Figure Lengend Snippet: Figure 4: The expressions and activations of death receptor proteins in P. trifoliata methanol extract (MEPT)‑treated MDA‑MB‑231 cells. Western blotting was performed for tumor necrosis factor receptor, TNFR type 1‑associated death domain, Fas, and FADD after administering MEPT at 0, 25, 50, or 100 μg/mL for 24 h, or 30 nM paclitaxel for 24 h (a), and expression levels were normalized using loading control (b)

Article Snippet: Primary antibodies for cleaved caspase 3, cleaved caspase 8, cleaved caspase 9, TNFR, TRADD, Fas, FADD, p38, phospho‐p38, JNK, and phospho‐JNK were purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Western Blot, Expressing, Control

Journal: Acta pharmacologica Sinica

Article Title: Ginsenoside Rb1 alleviates colitis in mice via activation of endoplasmic reticulum-resident E3 ubiquitin ligase Hrd1 signaling pathway.

doi: 10.1038/s41401-020-00561-9

Figure Lengend Snippet: Fig. 2 Effects of the ginsenoside Rb1 (GRb1) on inflammation and apoptosis in a DSS-induced mouse model of colitis. Expression levels of the colonic cytokines a MPO, b IL-1β, c IL-6, and d TNF-α were determined by ELISA. Western blot analysis of the inflammation-related proteins e iNOS, f COX2, and g p-NF-κB-p65 (calculated as p-NF-κB-p65/NF-κB-p65) and the apoptosis-related protein h cleaved-caspase 3 (cl-caspase 3), calculated as cl-caspase 3/caspase 3. Data are expressed as the mean ± SD. Values in the sham group were set to 100%, and other values are given relative to those in the sham group, **P < 0.01 compared with the sham group; ##P < 0.01 compared with the DSS control group; n = 3 samples for Western blot experiments; n = 6 samples for other experiments.

Article Snippet: Antibodies against Hrd1(13473-1-AP), CHOP(15204-1-AP), Fas (13098-1-AP), caspase 8(13423-1-AP), caspase 3 (19677-1-AP), cleaved-caspase 3 (cl-caspase 3) (19677-1-AP), and iNOS (14142-1-AP) were purchased from Proteintech Co., Ltd. (Wuhan, China).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Control

Journal: Acta pharmacologica Sinica

Article Title: Ginsenoside Rb1 alleviates colitis in mice via activation of endoplasmic reticulum-resident E3 ubiquitin ligase Hrd1 signaling pathway.

doi: 10.1038/s41401-020-00561-9

Figure Lengend Snippet: Fig. 3 Effects of the ginsenoside Rb1 (GRb1) on ER stress and Hrd1 in a DSS-induced mouse model of colitis. Western blot analysis of the ER stress marker proteins a GRP78, b PERK, c CHOP, and d caspase 12. e Western blot analysis of Hrd1. f Expression of Hrd1 determined by real- time PCR. g Immunohistochemical staining of Hrd1 in the colonic epithelium. Scale bars, 200, 100, and 50 μm. Data are expressed as the mean ± SD; **P < 0.01 compared with the sham group; ##P < 0.01 compared with the DSS control group; n = 3 samples for Western blot experiments; n = 6 samples for other experiments.

Article Snippet: Antibodies against Hrd1(13473-1-AP), CHOP(15204-1-AP), Fas (13098-1-AP), caspase 8(13423-1-AP), caspase 3 (19677-1-AP), cleaved-caspase 3 (cl-caspase 3) (19677-1-AP), and iNOS (14142-1-AP) were purchased from Proteintech Co., Ltd. (Wuhan, China).

Techniques: Western Blot, Marker, Expressing, Real-time Polymerase Chain Reaction, Immunohistochemical staining, Staining, Control

Journal: Acta pharmacologica Sinica

Article Title: Ginsenoside Rb1 alleviates colitis in mice via activation of endoplasmic reticulum-resident E3 ubiquitin ligase Hrd1 signaling pathway.

doi: 10.1038/s41401-020-00561-9

Figure Lengend Snippet: Fig. 5 Effect of the ginsenoside Rb1 (GRb1) on apoptosis. a Effect of GRb1 (200 μM) on IEC-6 cell apoptosis, as determined by flow cytometry. Effect of GRb1 on the expression of b Fas and c caspase 8 in vivo. Effect of GRb1 (200 μM) on the expression of Fas in LPS-treated IEC-6 cells in the absence (d) and presence (e) of siRNA targeting Hrd1. f Effects of GRb1 (200 μM) on IEC-6 cell apoptosis in the presence of siRNA targeting Hrd1. Data are expressed as the mean ± SD. **P < 0.01 compared with the corresponding control group; ##P < 0.01 compared with the DSS control group. n = 3 samples for Western blot experiments, n = 6 samples for other experiments.

Article Snippet: Antibodies against Hrd1(13473-1-AP), CHOP(15204-1-AP), Fas (13098-1-AP), caspase 8(13423-1-AP), caspase 3 (19677-1-AP), cleaved-caspase 3 (cl-caspase 3) (19677-1-AP), and iNOS (14142-1-AP) were purchased from Proteintech Co., Ltd. (Wuhan, China).

Techniques: Cytometry, Expressing, In Vivo, Control, Western Blot

Journal: Acta pharmacologica Sinica

Article Title: Ginsenoside Rb1 alleviates colitis in mice via activation of endoplasmic reticulum-resident E3 ubiquitin ligase Hrd1 signaling pathway.

doi: 10.1038/s41401-020-00561-9

Figure Lengend Snippet: Fig. 7 Effects of the ginsenoside Rb1 (GRb1) on Hrd1, ER stress, apoptosis, and inflammation in a TNBS-induced colitis model. Western blot analysis of a Hrd1, b CHOP, c GRP78, d cleaved-caspase 3 (cl-caspase 3) (calculated as cl-caspase 3/caspase 3), and e p-NF-κB-p65 (calculated as p-NF-κB-p65/NF-κB-p65). Data are expressed as the mean ± SD. **P < 0.01 compared with the sham group (n = 3 mice); ##P < 0.01 compared with the TNBS-induced colitis group; n = 3 mice.

Article Snippet: Antibodies against Hrd1(13473-1-AP), CHOP(15204-1-AP), Fas (13098-1-AP), caspase 8(13423-1-AP), caspase 3 (19677-1-AP), cleaved-caspase 3 (cl-caspase 3) (19677-1-AP), and iNOS (14142-1-AP) were purchased from Proteintech Co., Ltd. (Wuhan, China).

Techniques: Western Blot

Journal: Clinical and Translational Medicine

Article Title: NLK facilitates Caspase‐8 activation to drive macrophage PANoptosis in sepsis

doi: 10.1002/ctm2.70616

Figure Lengend Snippet: NLK deficiency disrupts PANoptosome assembly and augments RIPK1/3‐ dependent necrosome formation in vivo. (A, B) Representative immunoblots of FADD‐ and RIPK1‐associated complexes, together with corresponding input samples, in macrophages isolated from WT and NKO mice at 8 h post‐CLP. Co‐immunoprecipitates were probed for NLK, Caspase‐8, FADD, RIPK1, p‐RIPK1, RIPK3, and p‐RIPK3. (C–I) Quantification analysis of Caspase‐8, FADD, RIPK1, p‐RIPK1, RIPK3, and p‐RIPK3 in FADD‐associated complexes ( n = 3 independent biological replicates). (J–L) Quantification analysis of p‐RIPK1, RIPK3, and p‐RIPK3 in RIPK1‐associated complexes ( n = 3 independent biological replicates). (M) Representative confocal immunofluorescence images of lung macrophages stained for CD68 (green), Caspase‑8 (red), and ASC (cyan). Merged images indicate ASC–Caspase‐8 colocalisation; boxed regions show magnified views. Scale bars: 50 µm. Statistical significance was determined by using one‑way ANOVA with Bonferroni's post hoc test; * p < .05, ** p < .01 and ns indicates p > .05.

Article Snippet: Antibodies against CD68 (28058‐1‐AP), Caspase‐1 (22915‐1‐AP), p‐RIPK1 (66854‐1‐Ig), FADD ( P14906 ‐1‐AP), HA‐Tag (51064‐2‐AP, 66006‐2‐Ig), Flag‐Tag (66008‐4‐Ig, 20543‐1‐AP), and IgG (B900620) were purchased from Proteintech Group, Inc. (Wuhan, China).

Techniques: In Vivo, Western Blot, Isolation, Immunofluorescence, Staining

Journal: Clinical and Translational Medicine

Article Title: NLK facilitates Caspase‐8 activation to drive macrophage PANoptosis in sepsis

doi: 10.1002/ctm2.70616

Figure Lengend Snippet: NLK deficiency impairs PANoptosome assembly and enhances RIPK1/3‐dependent necrosome formation in macrophages. (A, B) Representative immunoblots of FADD‐ and RIPK1‐associated complexes, together with corresponding input samples, in macrophages isolated from WT and NKO mice at 3 h post‐LPS. Co‐IP were probed for NLK, Caspase‐8, FADD, RIPK1, p‐RIPK1, RIPK3, and p‐RIPK3. (C–I) Quantification analysis of Caspase‐8, FADD, RIPK1, p‐RIPK1, RIPK3, and p‐RIPK3 in FADD‐associated complexes ( n = 3 independent biological replicates). (J–L) Quantification analysis of p‐RIPK1, RIPK3, and p‐RIPK3 in RIPK1‐associated complexes ( n = 3 independent biological replicates). (M) Representative confocal immunofluorescence images showing the co‑localisation of RIPK3 (cyan), ASC (green), and Caspase‑8 (red) in PBS‐ or LPS‐treated BMDMs. Merged images indicate RIPK3–ASC–Caspase‐8 colocalisation; boxed regions show magnified views. Scale bars: 25 µm (merged), 10 µm (zoomed). Statistical differences were analysed by one‑way ANOVA with Bonferroni's post hoc test, * p < .05 and ** p < .01.

Article Snippet: Antibodies against CD68 (28058‐1‐AP), Caspase‐1 (22915‐1‐AP), p‐RIPK1 (66854‐1‐Ig), FADD ( P14906 ‐1‐AP), HA‐Tag (51064‐2‐AP, 66006‐2‐Ig), Flag‐Tag (66008‐4‐Ig, 20543‐1‐AP), and IgG (B900620) were purchased from Proteintech Group, Inc. (Wuhan, China).

Techniques: Western Blot, Isolation, Co-Immunoprecipitation Assay, Immunofluorescence

Journal: Journal of Biological Chemistry

Article Title: Fas-associated Protein with Death Domain (FADD)-independent Recruitment of c-FLIPL to Death Receptor 5

doi: 10.1074/jbc.m401056200

Figure Lengend Snippet: FIG. 3. DR5 and c-FLIPL interact in cell culture. A and B, interaction of c-FLIPL and DR5 proteins in co-immunoprecipitation studies. HEK 293 cells were co-transfected with expression vectors for vector alone, FLAG-tagged c-FLIPL, DR5, or c-FLIPL and DR5 as indicated. FLAG immunoprecipitates were analyzed for the presence of DR5 and DR5 immunoprecipitates were analyzed for the presence of c-FLIPL. The presence of DR5 and c-FLIP in cell extracts were verified by immunoblotting for DR5 or c-FLIP. C, interaction between c-FLIPp12 and DR5 in co- immunoprecipitation studies. HEK 293 cells were co-transfected with expression vectors for vector alone, FLAG-tagged c-FLIPp12, DR5, or c-FLIP12 and DR5 as indicated. FLAG immunoprecipitates were analyzed for the presence of DR5 and DR5 immunoprecipitates were analyzed for the presence of c-FLIPp12. The presence of DR5 in total cell lysate is shown by Western blot analysis with an anti-Myc antibody. D, interaction between endogenous c-FLIPL and endogenous DR5 is independent of FADD. Association of endogenous DR5 with endogenous c-FLIPL in parental Jurkat A3 and FADD-deficient Jurkat A3 cells is shown by immunoprecipitation with DR5 antibody followed by immunoblotting with c-FLIP antibody. The level of DR5 protein in the immunocomplex is determined by immunoblotting with DR5 antibody. An unrelated immune serum is used as a negative control. In the bottom panel, the level of FADD expression is shown for each cell line.

Article Snippet: The following antibodies were used in immunocomplex and DISC analysis: caspase-8 1C12 (Cell Signaling Technology, Beverly, MA), caspase-10 N-19 (Santa Cruz Biotechnology, Santa Cruz, CA), FADD H181 (Santa Cruz Biotechnology), FLIP AF821 (R&D Systems, Minneapolis, MN), FLIP G11 (Santa Cruz Biotechnology), and FLIP Dave 2 (Alexis Corporation).

Techniques: Cell Culture, Immunoprecipitation, Transfection, Expressing, Plasmid Preparation, Western Blot, Negative Control

Journal: Journal of Biological Chemistry

Article Title: Fas-associated Protein with Death Domain (FADD)-independent Recruitment of c-FLIPL to Death Receptor 5

doi: 10.1074/jbc.m401056200

Figure Lengend Snippet: FIG. 4. c-FLIPL interacts with DR5 in the absence of TRAIL stimulation and upon formation of the FADD/caspase-8 DISC, DR5, and c-FLIPL interaction is diminished. Time course of c- FLIPL and DR5 interaction and FADD/caspase-8 DISC formation was carried out in Jurkat cells that were treated with 10 ng/ml of TRAIL for 0, 5, 15, 30, 60, and 120 min. In stimulated cells, upon TRAIL treatment (10 ng/ml) for 1 h, cells were lysed and the assembled DISCs were immunoprecipitated with protein A and analyzed by Western blotting using antibodies to FADD, caspase-8, caspase-10, TRADD, c-FLIPL, c-FLIPp43, and c-FLIPs In unstimulated condition, cells were first lysed, and then TRAIL was added followed by analysis of the immunocomplex. In the absence of TRAIL treatment, c-FLIPL is associated with TRAIL receptor. Approximately, 5 min after TRAIL treatment FADD and caspase-8 are recruited to the DISC. At the same time, the level of c-FLIPL is diminished and the levels of c-FLIPp43 and c-FLIPS are increased in the complex.

Article Snippet: The following antibodies were used in immunocomplex and DISC analysis: caspase-8 1C12 (Cell Signaling Technology, Beverly, MA), caspase-10 N-19 (Santa Cruz Biotechnology, Santa Cruz, CA), FADD H181 (Santa Cruz Biotechnology), FLIP AF821 (R&D Systems, Minneapolis, MN), FLIP G11 (Santa Cruz Biotechnology), and FLIP Dave 2 (Alexis Corporation).

Techniques: Immunoprecipitation, Western Blot

Journal: Molecular Systems Biology

Article Title: An incoherent feedforward loop interprets NFκB/RelA dynamics to determine TNF‐induced necroptosis decisions

doi: 10.15252/msb.20209677

Figure Lengend Snippet:

Article Snippet: Rat FLIP Antibody [Dave‐2] , ProSci , Cat # XA‐1008.

Techniques: Recombinant, Sequencing, CRISPR, Control, cDNA Synthesis, SYBR Green Assay, Software, Microscopy, Imaging

Journal: Pharmaceuticals

Article Title: Carboxyl Group-Modified Myoglobin Induces TNF-α-Mediated Apoptosis in Leukemia Cells

doi: 10.3390/ph15091066

Figure Lengend Snippet: SEM-Mb-treated U937 cells underwent apoptosis. Without specific indication, U937 cells were treated with 1.5 μM SEM-Mb for 24 h. ( A ) SEM-Mb induced cell death in a concentration- and time-dependent manner. U937 cells were incubated with varying concentrations of SEM-Mb or Mb for 24 h. Cell viability was determined by MTT assay. Results are expressed as the percentage of cell survival relative to the control. Each value is the mean ± SD of triplicate determinations. (Inset) U937 cells were incubated with 1.5 μM SEM-Mb for indicated time periods. ( B ) Flow cytometry analyses of SEM-Mb-treated U937 cells using annexin V-FITC/propidium iodide double staining. (Left) Untreated control cells. (Right) U937 cells were treated with 1.5 μM SEM-Mb for 24 h. On flow cytometric scatter graphs, the left lower quadrant represents remaining live cells. The right lower quadrant represents the population of early apoptotic cells. The right upper quadrant represents the accumulation of late apoptotic cells. ( C ) Western blot analyses showing the degradation of procaspase-3/-8 and PARP in SEM-Mb-treated cells. ( D ) Pretreatment with caspase inhibitors restored the viability of SEM-Mb -treated cells. U937 cells were pretreated with 10 μM Z-DEVD-FMK (caspase-3 inhibitor) or Z-IETD-FMK (caspase-8 inhibitor) for 1 h, and then incubated with 1.5 μM SEM-Mb for 24 h. Cell viability was determined by MTT assay. The values represent averages of three independent experiments with triplicate measurement (mean ± SD, * p < 0.05). ( E ) Effect of SEM-Mb on the production of t-Bid and the expression of BCL2 family proteins. ( F ) Dissipation of mitochondrial membrane potential (∆Ψm) in SEM-Mb-treated cells. U937 cells were treated with 1.5 μM SEM-Mb for 24 h, and then incubated with rhodamine 123 for 15 min. The loss of ∆Ψm was analyzed by flow cytometry. ( G ) Transfection of FADD siRNA inhibited SEM-Mb-induced the degradation of procaspase-8/-3 and the production of t-Bid. U937 cells were transfected with 100 nM control siRNA or FADD siRNA, respectively. After 24 h post-transfection, the cells were treated with 1.5 μM SEM-Mb for 24 h. ( H ) Depletion of FADD inhibited SEM-Mb-induced cell death (mean ± SD, * p < 0.05).

Article Snippet: Gene silencing experiments were performed using FADD siRNA, NOX4 siRNA, and negative control siRNA (Santa Cruz Biotechnology).

Techniques: Concentration Assay, Incubation, MTT Assay, Control, Flow Cytometry, Double Staining, Western Blot, Expressing, Membrane, Transfection

Journal: Pharmaceuticals

Article Title: Carboxyl Group-Modified Myoglobin Induces TNF-α-Mediated Apoptosis in Leukemia Cells

doi: 10.3390/ph15091066

Figure Lengend Snippet: NOX4-mediated SIRT3 degradation regulated p38 MAPK/TTP axis-dependent TNF-α upregulation in SEM-Mb-treated cells. U937 cells were directly treated with 1.5 μM SEM-Mb for 24 h or pre-treated with 1 μM MG132 or 10 μM GLX351322 for 1 h and then incubated with 1.5 μM SEM-Mb for 24 h. ( A ) Effect of SEM-Mb on SIRT3 expression. ( B ) qRT-PCR analyses of SIRT3 mRNA levels in SEM-Mb-treated cells (mean ± SD, NS, not statistically significant). Effect of ( C ) MG132 and ( D ) GLX351322 on the SIRT3 expression in SEM-Mb-treated cells. ( E ) Effect of NOX4 depletion on SIRT3 expression in U937 cells. U937 cells were transfected with 100 nM negative control siRNA or NOX4 siRNA, respectively. After 24 h post-transfection, the cells were treated with 1.5 μM SEM-Mb for 24 h. ( F ) Effect of SIRT3 overexpression on SEM-Mb-induced the production of mitochondrial ROS in U937 cells (mean ± SD, * p < 0.05). U937 cells were transfected with empty vector or pCMV3-N-His-SIRT3, respectively. After 24 h post-transfection, the transfected cells were treated with 1.5 μM SEM-Mb for 24 h. ( G ) Effect of SIRT3 overexpression on the viability of SEM-Mb-treated cells (mean ± SD, * p < 0.05). ( H ) Effect of SIRT3 overexpression on the expression of p-p38 MAPK, TTP, and TNF-α in SEM-Mb-treated U937 cells.

Article Snippet: Gene silencing experiments were performed using FADD siRNA, NOX4 siRNA, and negative control siRNA (Santa Cruz Biotechnology).

Techniques: Incubation, Expressing, Quantitative RT-PCR, Transfection, Negative Control, Over Expression, Plasmid Preparation

Journal: Journal of Diabetes Investigation

Article Title: Kbtbd11 gene expression in adipose tissue increases in response to feeding and affects adipocyte differentiation

doi: 10.1111/jdi.12995

Figure Lengend Snippet: Kbtbd11 expression levels in epididymal white adipose tissue. (a) The expression of Kbtbd11 in epididymal white adipose tissue of C57BL/6J mice; mice were fasted for 24 h or fasted for 24 h/re‐fed for 12 h; n = 3 per group, * P < 0.01 versus fasted. (b) The expression of Kbtbd11 in epididymal white adipose tissue of diet‐induced obesity mice; diet‐induced obesity mice were fed a high‐fat diet for 1 month. The mice were fasted for 24 h; n = 3 per group, * P < 0.01 versus chow.

Article Snippet: Furthermore, Kbtbd11 knockdown inhibited adipogenesis, but only before MCE (not mature 3T3‐L1 adipocytes), and Kbtbd11 overexpression induced MCE, leading to the expression of Cebpa and Pparg (Figure c,f).

Techniques: Expressing

Journal: Journal of Diabetes Investigation

Article Title: Kbtbd11 gene expression in adipose tissue increases in response to feeding and affects adipocyte differentiation

doi: 10.1111/jdi.12995

Figure Lengend Snippet: Kbtbd11 messenger ribonucleic acid (mRNA) expression in differentiating 3T3‐L1 cells. Total RNA was extracted from 3T3‐L1 cells at day 0, 2, 4, 8 and 10 after the induction of differentiation by treatment with an adipogenic cocktail; n = 3 per group, * P < 0.01 versus day 0.

Article Snippet: Furthermore, Kbtbd11 knockdown inhibited adipogenesis, but only before MCE (not mature 3T3‐L1 adipocytes), and Kbtbd11 overexpression induced MCE, leading to the expression of Cebpa and Pparg (Figure c,f).

Techniques: Expressing

Journal: Journal of Diabetes Investigation

Article Title: Kbtbd11 gene expression in adipose tissue increases in response to feeding and affects adipocyte differentiation

doi: 10.1111/jdi.12995

Figure Lengend Snippet: Kbtbd11 messenger ribonucleic acid (mRNA) expression in response to adipogenic stimuli at early time points. 3T3‐L1 preadipocytes were cultured to confluence, serum‐starved for 12 h and treated with (a) 5 μg/mL insulin, (b) 2.5 μmol/L dexamethasone (DEX) or (c) 200 μmol/L 3‐isobutyl‐1‐methylxanthine (IBMX) for the indicated times; n = 3 per group, * P < 0.01 versus 0 h.

Article Snippet: Furthermore, Kbtbd11 knockdown inhibited adipogenesis, but only before MCE (not mature 3T3‐L1 adipocytes), and Kbtbd11 overexpression induced MCE, leading to the expression of Cebpa and Pparg (Figure c,f).

Techniques: Expressing, Cell Culture

Journal: Journal of Diabetes Investigation

Article Title: Kbtbd11 gene expression in adipose tissue increases in response to feeding and affects adipocyte differentiation

doi: 10.1111/jdi.12995

Figure Lengend Snippet: The effects of knockdown and overexpression of Kbtbd11 on 3T3‐L1 cellular differentiation. (a) The expression of Kbtbd11 messenger ribonucleic acid (mRNA) in Kbtbd11 knockdown 3T3‐L1 cells at day 8. Kbtbd11 knockdown adenovirus particles were used with either of the two independent Kbtbd11 short hairpin ribonucleic acid (shRNA) constructs (shRNA#1 and shRNA#2); n = 3 per group, * P < 0.01 versus LacZ‐specific short hairpin ribonucleic acid (shLacZ); (b) triglyceride accumulation in 3T3‐L1 cells on day 8, visualized using Oil Red O staining; (c) the mRNA levels in 3T3‐L1 cells expressing each shRNA at various time points after inducing differentiation; n = 3 per group, * P < 0.01 versus shLacZ; (d) the expression of Kbtbd11 mRNA in Kbtbd11 ‐overexpressing 3T3‐L1 cells at day 8. Cells were infected with adenoviral vectors for expressing green fluorescent protein (GFP) or mouse Kbtbd11 ; n = 3 per group, * P < 0.01 versus GFP; (e) triglyceride accumulation in 3T3‐L1 cells on day 6 was detected using Oil Red O staining. (f) Relative mRNA levels in each group of 3T3‐L1 cells at various time points after inducing differentiation; n = 3 per group, * P < 0.01 versus GFP.

Article Snippet: Furthermore, Kbtbd11 knockdown inhibited adipogenesis, but only before MCE (not mature 3T3‐L1 adipocytes), and Kbtbd11 overexpression induced MCE, leading to the expression of Cebpa and Pparg (Figure c,f).

Techniques: Knockdown, Over Expression, Cell Differentiation, Expressing, shRNA, Construct, Staining, Infection

Journal: Journal of Diabetes Investigation

Article Title: Kbtbd11 gene expression in adipose tissue increases in response to feeding and affects adipocyte differentiation

doi: 10.1111/jdi.12995

Figure Lengend Snippet: Effects of knockdown of Kbtbd11 on mature 3T3‐L1 adipocytes. (a) Triglyceride accumulation in Kbtbd11 ‐knockdown mature 3T3‐L1 adipocytes at 48 h after either of the two independent Kbtbd11 short hairpin ribonucleic acid (shRNA) adenoviral infections (shRNA#1 and shRNA#2) visualized using Oil Red O staining; (b) RNA was harvested at 48 h after adenoviral infection, and expression levels of Kbtbd11 , adipocytes ( Pparg and aP2 ) and inflammation markers ( Tnfa and Il6 ), and lipogenic ( Fasn ) and proapoptotic genes ( Bax and Bcl2 ) were measured using quantitative polymerase chain reaction; n = 3 per group, * P < 0.01 versus shLacZ.

Article Snippet: Furthermore, Kbtbd11 knockdown inhibited adipogenesis, but only before MCE (not mature 3T3‐L1 adipocytes), and Kbtbd11 overexpression induced MCE, leading to the expression of Cebpa and Pparg (Figure c,f).

Techniques: Knockdown, shRNA, Staining, Infection, Expressing, Real-time Polymerase Chain Reaction