Journal: Stem Cells International

Article Title: Electroacupuncture Improved Hippocampal Neurogenesis following Traumatic Brain Injury in Mice through Inhibition of TLR4 Signaling Pathway

doi: 10.1155/2017/5841814

Figure Lengend Snippet: EA treatment exerted an inhibitory effect on both of the two downstream pathways of TLR4 in the hippocampus posttrauma. Hippocampal TLR4, Myd88, TRAF6, TRAM, TRIF, and NF- κ B expression of the sham, TBI, TBI + EA, TBI + EA + LPS, TBI + EA + Veh, and sham + EA groups ( n = 3 in each group) were detected by WB at 35 days after TBI. (a–f) The six kinds of proteins in TLR4/Myd88 κ B and TLR4/TRIF/ κ B signaling pathways were upregualted in the TBI group compared with the sham group and the sham + EA group. EA treatment resulted in a significant decrease in the TBI + EA group, but the effects were attenuated by LPS administration in the TBI + EA + LPS group. No difference of the proteins between the TBI + EA and TBI + EA + Veh groups was observed. ∗ P < 0.05 versus the TBI group, † P < 0.05 versus the TBI + EA + LPS group.

Article Snippet: Membranes were blocked with 5% nonfat milk solution in tris-buffered saline with 0.1% Triton X-100 (TBST) for 1 hour and then incubated overnight at 4°C with appropriate primary antibodies as below: rabbit anti-mouse TLR4 antibody (1 : 1000, Thermo Fisher, PA5-23125, Rockford, IL, USA), rabbit anti-mouse myeloid differentiation factor 88 (Myd88) antibody (1 : 500, Santa Cruz, sc-17320, Dallas, TX, USA), rabbit anti-mouse TNFR-associated factor (TRAF6) antibody (1 : 1000, Novus Biological, NB100-56179, Littleton, CO, USA), rabbit anti-mouse toll/IL-1 receptor domain-containing adapter-induced interferon- β (TRIF) antibody (1 : 1000, Enzo Life Sciences, ALX-215-016, Farmingdale, NY, USA), rabbit anti-mouse TRIF-related adaptor molecule (TRAM) antibody (1 : 1000, OriGene, TA-306163, Rockville, MD, USA), rabbit anti-mouse nuclear factor- κ B (NF- κ B) p65 antibody (1 : 1000, GeneTex, GTX21893, Irvine, CA, USA), and rabbit anti- β -actin antibody (1 : 2000, Proteintech, 20536-1-AP, Rosemont, IL, USA).

Techniques: Expressing

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: Decitabine promotes degradation of DNMT1 and EZH2 via the ubiquitination pathway and inhibits colorectal cancer progression

doi: 10.1007/s13402-025-01136-8

Figure Lengend Snippet: TRAF6 mediates DAC-induced DNMT1 ubiquitination and degradation. (A) Reduced TRAF6 mRNA expression in colorectal cancer tissues compared to adjacent normal tissues (TCGA dataset; RNA-seq analysis; **** p < 0.0001). (B) Kaplan-Meier analysis reveals prolonged post-progression survival (PPS) in patients with high TRAF6 expression versus low expression (log-rank test, p = 0.0061). (C) Inverse correlation between DNMT1 and TRAF6 mRNA expression in TCGA colorectal cohorts (Spearman’s r = -0.1603, p = 0.0008061). (D) Immunofluorescence showing co-localization of DNMT1 (green) and TRAF6 (red) in HCT8 and SW620 (DAPI: blue; scale bar:100 μm). (E) TRAF6 protein upregulation in DAC-treated cells. (F) TRAF6 knockdown (siTRAF6) significantly elevated DNMT1 protein levels. (G) IP-Western blot showing reduced DNMT1 ubiquitination upon TRAF6 knockdown (anti-Ub antibody; IgG as IP negative control). (H) TRAF6 knockdown blocks DAC-induced DNMT1 degradation I , J. TRAF6 protein levels inversely correlate with DNMT1 modulation. K-M. qRT-PCR reveals TRAF6 transcriptional upregulation following DNMT1 knockdown (siDNMT1) or DAC treatment (500nM, 48 h), and suppression by DNMT1 OE(* p < 0.05,** p < 0.01, *** p < 0.001, **** p < 0.0001). N. DNMT1 induces hypermethylation of the TRAF6 promoter. Methylation-specific PCR (MSP) using methylated (M) and unmethylated (U) primers. DNMT1 OE enhances methylation (intensified M-band). O. Dual-luciferase assays show DNMT1 knockdown or treated with DAC increases TRAF6 promotor activity in HEK293T cells(** p < 0.01)

Article Snippet: Cells were washed three in PBS and blocked for 1 h with 5% BSA in PBS and then incubated with primary antibodies DNMT1 (abcam, ab188453) and TRAF6 (santa cruz, sc-8409) at 4 °C overnight.

Techniques: Ubiquitin Proteomics, Expressing, RNA Sequencing, Immunofluorescence, Knockdown, Western Blot, Negative Control, Quantitative RT-PCR, Methylation, Luciferase, Activity Assay

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: Decitabine promotes degradation of DNMT1 and EZH2 via the ubiquitination pathway and inhibits colorectal cancer progression

doi: 10.1007/s13402-025-01136-8

Figure Lengend Snippet: DNMT1 silences TRAF6 to block EZH2 ubiquitination. A-C. DNMT1 modulation regulates EZH2 protein levels in CRC cells: Western blot analysis of EZH2 in DNMT1-overexpressing and DNMT1-knockdown( A , B ), DAC-treated ( C ) cells. D-G. qRT-PCR analysis of EZH2 mRNA in HCT8 ( D ) and SW480 ( E ) cells with DNMT1 modulation (ns: not significant). H-I. Cycloheximide (CHX, 50 µg/mL) chase assay showing reduced EZH2 half-life in DNMT1-knockdown HCT8 cells. J. Proteasome inhibitor MG132 (10 µM, 6 h) rescues EZH2 protein levels in DNMT1-depleted cells. K-L. Endogenous ubiquitination assays showing increased polyubiquitinated EZH2 in DNMT1-knockdown or treated with DAC cells (IP with anti-EZH2 followed by anti-Ub Western blot; input: 5% lysate). M. TCGA data analysis reveals significantly elevated EZH2 mRNA expression in colorectal cancer (CRC) tissues compared to adjacent normal tissues. N. Expression correlation analysis demonstrates a significant inverse relationship between TRAF6 and EZH2 mRNA levels in colorectal cohorts ( p < 0.0001). O , P. Overexpression of TRAF6 enhances polyubiquitination of EZH2, as detected by ubiquitin (Ub) immunoblotting in CRC cells. TRAF6 knockdown reduces EZH2 ubiquitination levels. Lysates were immunoprecipitated (IP) with anti-EZH2 and immunoblotted (IB) with anti-Ub. Q. Co-IP assay demonstrating strengthened EZH2-TRAF6 interaction in DNMT1-depleted cells (lysates immunoprecipitated with anti-EZH2, blotted for TRAF6; input: 5% lysate). R. Dual inhibition of TRAF6 and DNMT1 rescues EZH2 hyperubiquitination (IP-Western blot with siTRAF6/siDNMT1 co-treatment

Article Snippet: Cells were washed three in PBS and blocked for 1 h with 5% BSA in PBS and then incubated with primary antibodies DNMT1 (abcam, ab188453) and TRAF6 (santa cruz, sc-8409) at 4 °C overnight.

Techniques: Blocking Assay, Ubiquitin Proteomics, Western Blot, Knockdown, Quantitative RT-PCR, Expressing, Over Expression, Immunoprecipitation, Co-Immunoprecipitation Assay, Inhibition

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: Decitabine promotes degradation of DNMT1 and EZH2 via the ubiquitination pathway and inhibits colorectal cancer progression

doi: 10.1007/s13402-025-01136-8

Figure Lengend Snippet: EZH2 is a downstream effector of DNMT1 in CRC progression. (A) Western blot analysis showing that EZH2 knockdown reverses DNMT1-induced upregulation of cyclinD1 and cyclinE2 in HCT8 and SW480 cells. (B) Colony formation assays demonstrating that EZH2 depletion abolishes DNMT1-driven proliferation in HCT8 cells (** p < 0.01 vs. DNMT1-OE + siControl). C-D. Colony formation assays demonstrating that TRAF6 overexpression abolishes DNMT1/EZH2-driven proliferation in HCT8 cells (** p < 0.01 vs. DNMT1-OE + Vector, ** p < 0.01 vs. EZH2-OE + Vector). E. Transwell migration assays showing suppression of DNMT1-enhanced motility by EZH2 knockdown in HCT8 cells(*** p < 0.001 vs. DNMT1-OE + siControl); Matrigel invasion assays confirming EZH2-dependent regulation of DNMT1-induced invasiveness in SW480 cells(** p < 0.01 vs. DNMT1-OE + siControl; scale bar: 100 μm). F. Transwell migration assays showing suppression of DNMT1-enhanced motility by TRFA6 overexpression in HCT8 cells(** p < 0.01 vs. DNMT1-OE + Vector); Matrigel invasion assays confirming TRAF6-dependent regulation of DNMT1-induced invasiveness in HCT8 cells(*** p < 0.001 vs. DNMT1-OE + Vector; scale bar: 100 μm). G. Transwell migration assays showing suppression of EZH2-enhanced motility by TRFA6 overexpression in HCT8 cells(*** p < 0.001 vs. EZH2-OE + Vector); Matrigel invasion assays confirming TRAF6-dependent regulation of EZH2-induced invasiveness in HCT8 cells(*** p < 0.001 vs. EZH2-OE + Vector; scale bar: 100 μm)

Article Snippet: Cells were washed three in PBS and blocked for 1 h with 5% BSA in PBS and then incubated with primary antibodies DNMT1 (abcam, ab188453) and TRAF6 (santa cruz, sc-8409) at 4 °C overnight.

Techniques: Western Blot, Knockdown, Over Expression, Plasmid Preparation, Migration

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: Decitabine promotes degradation of DNMT1 and EZH2 via the ubiquitination pathway and inhibits colorectal cancer progression

doi: 10.1007/s13402-025-01136-8

Figure Lengend Snippet: Effects of Decitabine (DAC) Alone and in Combination with Tazemetostat (TAZ) on the Biological Behaviors of SW480 Cells and Analysis of Related Molecular Expressions. (A) Representative images of colonies formed by SW480 cells treated with DMSO (vehicle control), decitabine (DAC, 500 nM) alone, tazemetostat (TAZ, 1 µM) alone, or the combination (DAC + TAZ) (*** p < 0.001). (B) Representative images (left) and quantitative analysis (right) of the migration (upper panel) and invasion (lower panel) of SW480 cells after treatment with DMSO (vehicle control), decitabine (DAC, 500 nM) alone, tazemetostat (TAZ, 1 µM) alone, or the combination (DAC + TAZ) (** p < 0.01, *** p < 0.001). (C) The expression levels of DNMT1, EZH2 and TRAF6 in colorectal cancer tissues and adjacent non-tumor tissues. (D) Regulatory Relationships among Decitabine, TRAF6, DNMT1 and EZH2

Article Snippet: Cells were washed three in PBS and blocked for 1 h with 5% BSA in PBS and then incubated with primary antibodies DNMT1 (abcam, ab188453) and TRAF6 (santa cruz, sc-8409) at 4 °C overnight.

Techniques: Control, Migration, Expressing

Journal: Journal of experimental & clinical cancer research : CR

Article Title: Cinchonine induces apoptosis of HeLa and A549 cells through targeting TRAF6.

doi: 10.1186/s13046-017-0502-8

Figure Lengend Snippet: Fig. 1 Docking result of cinchonine. a Binding of cinchonine (in cyan) with TRAF6. b The -OH group of cinchonine forges one hydrogen bond (in pink) with residues Asp57 in TRAF6. c The binding of Ubc13 (in red) to TRAF6 (in blue). d The influence of cinchonine on the interaction between TRAF6 and Ubc13

Article Snippet: Antibodies used in this study include the following: Ubiquitination antibody and rabbit anti-TRAF6 polyclonal antibody (Santa Cruz, USA); Rabbit polyclonal antibody against total AKT, phosphorylation-AKT (Thr-308), phosphorylation-AKT (Ser-473), total TAK1, phosphorylation-TAK1 (Thr-184/Thr-187), Bax, and Bcl-2 (Cell Signaling Technology, USA); Rabbit polyclonal antibody against TRAF6, ALEXA FLUOR 647 conjugated (Bioss Inc. USA); Rabbit anti β-actin polyclonal antibody, Goat anti-mouse, and goat anti-rabbit IgG polyclonal antibody (Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd, China).

Techniques: Binding Assay

Journal: Journal of experimental & clinical cancer research : CR

Article Title: Cinchonine induces apoptosis of HeLa and A549 cells through targeting TRAF6.

doi: 10.1186/s13046-017-0502-8

Figure Lengend Snippet: Fig. 8 Visualizing binding of cinchonine with TRAF6 in HeLa and A549 cells. a Synthesis of a fluorescent probe conjugated cinchonine. b Immunofluorescence staining images of the localizations of TRAF6 and cinchonine in cells. Modified cinchonine (green fluorescence) and TRAF6 (red fluorescence) were co-located to cytoplasm

Article Snippet: Antibodies used in this study include the following: Ubiquitination antibody and rabbit anti-TRAF6 polyclonal antibody (Santa Cruz, USA); Rabbit polyclonal antibody against total AKT, phosphorylation-AKT (Thr-308), phosphorylation-AKT (Ser-473), total TAK1, phosphorylation-TAK1 (Thr-184/Thr-187), Bax, and Bcl-2 (Cell Signaling Technology, USA); Rabbit polyclonal antibody against TRAF6, ALEXA FLUOR 647 conjugated (Bioss Inc. USA); Rabbit anti β-actin polyclonal antibody, Goat anti-mouse, and goat anti-rabbit IgG polyclonal antibody (Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd, China).

Techniques: Binding Assay, Immunofluorescence, Staining, Modification, Fluorescence

Journal: Cellular signalling

Article Title: IRAK2 overexpression restrains prostate cancer progression by regulation of TRAF6 ubiquitination.

doi: 10.1016/j.cellsig.2024.111508

Figure Lengend Snippet: Fig. 3. IRAK2 interacted with TRAF6 in prostate cancer cells. (A) The interaction of IRAK2 was detected on STRING database. (B) Interaction between IRAK2 and TRAF6 was assessed by CO-IP. (C) Detection of TRAF6 expression in PC-3 cells transfected IRAK2-overexpression plasmid using Western blot. (D) TRAF6 level was detected in DU145 cells after transfection of IRAK2-overexpression plasmid by Western blot. (E) Detection of NF-κB expression in PC-3 cells transfected IRAK2- overexpression plasmid using Western blot. (F) NF-κB level was detected in DU145 cells after transfection of IRAK2-overexpression plasmid by Western blot. All data were presented as the mean ± SD from at least three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, compared with the NC group.

Article Snippet: Overnight treatment with primary antibodies GAPDH (5174S, CST, US), IRAK2 (human, ab192249, Abcam, US), IRAK2 (mice, ab62419, Abcam, US), TRAF6 (human, AF3284, RD systems, US), TRAF6 (mice, ab33915, Abcam, US), anti-ubiquitin (ab7254, Abcam, US) was conducted at 4 ◦C.

Techniques: Co-Immunoprecipitation Assay, Expressing, Transfection, Over Expression, Plasmid Preparation, Western Blot

Journal: Cellular signalling

Article Title: IRAK2 overexpression restrains prostate cancer progression by regulation of TRAF6 ubiquitination.

doi: 10.1016/j.cellsig.2024.111508

Figure Lengend Snippet: Fig. 4. IRAK2 inhibited ubiquitination and degradation of TRAF6 in prostate cancer cells. (A) TRAF6 expression was assessed in PC-3 or IRAK2-overexpression PC-3 cells after treatment with 50 mg/mL cycloheximide (CHX) for 0 h, 2 h, 4 h and 8 h by Western blot. (B) TRAF6 expression was assessed in DU145 or IRAK2- overexpression DU145 cells after treatment with 50 mg/mL cycloheximide (CHX) for 0 h, 2 h, 4 h and 8 h by Western blot. (C) TRAF6 expression was detected in PC-3 and IRAK2-overexpression PC-3 cells after treatment with 10 μm MG132 for 0 h and 8 h by Western blot. (D) TRAF6 expression was detected in DU145 and IRAK2-overexpression DU145 cells after treatment with 10 μm MG132 for 0 h and 8 h by Western blot. (E) Western blot was assessed to detection of ubiquitination level in IRAK2-overexpression PC-3 cells. (F) Ubiquitination level was assessed in IRAK2-overexpression DU145 cells by Western blot. (G) Detection of FLAG level in PC-3 cells co-transfected HA-IRAK2 (IRAK2 overexpression), FLAG-TRAF6 (TRAF6 overexpression) and MYC-Ub (Ubiquitin overexpression) plasmids using IP assay. (H) Myc level was detected in PC-3 cells after transfection of HA-IRAK2 (IRAK2 overexpression), FLAG-TRAF6 (TRAF6 overexpression) and MYC-Ub (Ubiquitin overexpression) plasmids by IP assay. (I) FLAG level was detected in DU145 cells after transfection of HA-IRAK2 (IRAK2 overexpression), FLAG-TRAF6 (TRAF6 overexpression) and MYC-Ub (Ubiquitin overexpression) plasmids by IP assay. (J) Myc level was detected in DU145 cells after transfection of HA-IRAK2 (IRAK2 overexpression), FLAG-TRAF6 (TRAF6 overexpression) and MYC-Ub (Ubiquitin overexpression) plasmids by IP assay. All data were presented as the mean ± SD from at least three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, compared with the NC group.

Article Snippet: Overnight treatment with primary antibodies GAPDH (5174S, CST, US), IRAK2 (human, ab192249, Abcam, US), IRAK2 (mice, ab62419, Abcam, US), TRAF6 (human, AF3284, RD systems, US), TRAF6 (mice, ab33915, Abcam, US), anti-ubiquitin (ab7254, Abcam, US) was conducted at 4 ◦C.

Techniques: Ubiquitin Proteomics, Expressing, Over Expression, Western Blot, Transfection

Journal: Cellular signalling

Article Title: IRAK2 overexpression restrains prostate cancer progression by regulation of TRAF6 ubiquitination.

doi: 10.1016/j.cellsig.2024.111508

Figure Lengend Snippet: Fig. 5. IRAK2 affected prostate cancer progression by regulating TRAF6 expression. (A) Detection of TRAF6 expression in IRAK2-overexpression PC-3 cells trans fected the TRAF6-sh plasmid using Western blot. (B) Detection of TRAF6 expression in IRAK2-overexpression DU145 cells transfected the TRAF6-sh plasmid using Western blot. (C) Cell viability after transfection of the TRAF6-sh plasmid was analyzed by MTT assay in IRAK2-overexpression PC-3 cells. (D) Cell viability after transfection of the TRAF6-sh plasmid was analyzed by MTT assay in IRAK2-overexpression DU145 cells. (E) Transwell assay was conducted to detect the migratory and invasive capacities of PC-3 cells transfected the TRAF6-sh plasmid in IRAK2-overexpression PC-3 cells (scare bar: 400 μm). (F) Transwell assay was conducted to detect the migratory and invasive capacities of DU145 cells transfected the TRAF6-sh plasmid in IRAK2-overexpression DU145 cells (scare bar: 400 μm). (G) Detection of migratory capacity in IRAK2-overexpression PC-3 cells transfected the TRAF6-sh plasmid using wound-healing migration assay (scare bar: 200 μm). (H) Detection of migratory capacity in IRAK2-overexpression DU145 cells transfected the TRAF6-sh plasmid using wound-healing migration assay (scare bar: 200 μm). All data were presented as the mean ± SD from at least three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, compared with the NC group.

Article Snippet: Overnight treatment with primary antibodies GAPDH (5174S, CST, US), IRAK2 (human, ab192249, Abcam, US), IRAK2 (mice, ab62419, Abcam, US), TRAF6 (human, AF3284, RD systems, US), TRAF6 (mice, ab33915, Abcam, US), anti-ubiquitin (ab7254, Abcam, US) was conducted at 4 ◦C.

Techniques: Expressing, Over Expression, Plasmid Preparation, Western Blot, Transfection, MTT Assay, Transwell Assay, Migration

Journal: Cellular signalling

Article Title: IRAK2 overexpression restrains prostate cancer progression by regulation of TRAF6 ubiquitination.

doi: 10.1016/j.cellsig.2024.111508

Figure Lengend Snippet: Fig. 6. TRAF6 E3 ligase activity did not participate in the regulation of prostate cancer progression. (A) Ubiquitination level in 293 T cells transfected with TRAF6 overexpression or C70ATRAF6 overexpression plasmids was analyzed by Western blot. (B) Detection of IRAK2 and C70ATRAF6 level in 293 T cells co-transfected with IRAK2-overexpression and C70ATRAF6-expression plasmids using Western blot. (C) IP assay was used to detect FLAG level in PC-3 cells after transfection of FLAG- TRAF6 plasmid, FLAG-C70ATRAF6 plasmid, HA-IRAK2 and FLAG-TRAF6 plasmid and HA-IRAK2 and FLAG-C70ATRAF6 plasmid, respectively. (D) IP assay was used to detect FLAG level in DU145 cells after transfection of FLAG-TRAF6 plasmid, FLAG-C70ATRAF6 plasmid, HA-IRAK2 and FLAG-TRAF6 plasmid and HA-IRAK2 and FLAG-C70ATRAF6 plasmid, respectively. (E) The migratory capacity of PC-3 or DU145cells was detected by Transwell assay after transfection of TRAF6- overexpression plasmid or C70ATRAF6-overexpression plasmid, respectively (scare bar: 200 μm). (F) The migratory capacity of DU145cells was detected by Trans well assay after transfection of TRAF6-overexpression plasmid or C70ATRAF6-overexpression plasmid, respectively (scare bar: 200 μm). (G) Cell viability after transfection of the TRAF6-overexpression plasmid or C70ATRAF6-overexpression plasmid was analyzed by MTT assay in PC-3 cells. (H) Cell viability after transfection of the TRAF6-overexpression plasmid or C70ATRAF6-overexpression plasmid was analyzed by MTT assay in DU145 cells. All data were presented as the mean ± SD from at least three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, compared with the NC group.

Article Snippet: Overnight treatment with primary antibodies GAPDH (5174S, CST, US), IRAK2 (human, ab192249, Abcam, US), IRAK2 (mice, ab62419, Abcam, US), TRAF6 (human, AF3284, RD systems, US), TRAF6 (mice, ab33915, Abcam, US), anti-ubiquitin (ab7254, Abcam, US) was conducted at 4 ◦C.

Techniques: Activity Assay, Ubiquitin Proteomics, Transfection, Over Expression, Western Blot, Expressing, Plasmid Preparation, Transwell Assay, MTT Assay

Journal: Cellular signalling

Article Title: IRAK2 overexpression restrains prostate cancer progression by regulation of TRAF6 ubiquitination.

doi: 10.1016/j.cellsig.2024.111508

Figure Lengend Snippet: Fig. 8. IRAK2 inhibited the progression of prostate cancer in vivo. (A) BALB/c nude mice in the NC group and IRAK2-OE group (n = 10), were dissected after subcutaneous injection 9 days. (B) The mice weight was measured with calipers every 3 days. (C) The mice tumor volume was measured with calipers every 3 days, and volume was calculated as (width2 × length)/2. (D) The mRNA level of IRAK2 and TRAF6 were assessed using qRT-PCR. (E) Western blotting detected the expression of IRAK2 and TRAF6 in the NC group and IRAK2-OE group. All data were presented as the mean ± SD from at least three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, compared with the NC group.

Article Snippet: Overnight treatment with primary antibodies GAPDH (5174S, CST, US), IRAK2 (human, ab192249, Abcam, US), IRAK2 (mice, ab62419, Abcam, US), TRAF6 (human, AF3284, RD systems, US), TRAF6 (mice, ab33915, Abcam, US), anti-ubiquitin (ab7254, Abcam, US) was conducted at 4 ◦C.

Techniques: In Vivo, Injection, Quantitative RT-PCR, Western Blot, Expressing

Journal: Cellular signalling

Article Title: IRAK2 overexpression restrains prostate cancer progression by regulation of TRAF6 ubiquitination.

doi: 10.1016/j.cellsig.2024.111508

Figure Lengend Snippet: Fig. 9. A schematic diagram of IRAK2 regulating TRAF6 expression in prostate cancer. First, IRAK2 decreased the ubiquitination level of TRAF6 by interacting with TRAF6, thereby increasing the expression of TRAF6. Next, up-regulation of TRAF6 induced the inhibition of prostate cancer.

Article Snippet: Overnight treatment with primary antibodies GAPDH (5174S, CST, US), IRAK2 (human, ab192249, Abcam, US), IRAK2 (mice, ab62419, Abcam, US), TRAF6 (human, AF3284, RD systems, US), TRAF6 (mice, ab33915, Abcam, US), anti-ubiquitin (ab7254, Abcam, US) was conducted at 4 ◦C.

Techniques: Expressing, Ubiquitin Proteomics, Inhibition