Journal: International Dental Journal

Article Title: C1QBP Drives M2 Macrophage Polarization Via TRAF2-CCL2 to Promote Oral Squamous Cell Carcinoma Progression

doi: 10.1016/j.identj.2025.103938

Figure Lengend Snippet: Knockdown of C1QBP upregulates the expression of key genes TRAF2 and CCL2 in the TNF signaling pathway. (A) qRT-PCR detection of significantly increased mRNA levels of TRAF2 and CCL2 in C1QBP-knockdown OSCC cells ( P < .05). (B) WB demonstration of significantly upregulated TRAF2 and CCL2 protein levels in C1QBP-knockdown OSCC cells. (C) Quantitative analysis of WB experimental data.

Article Snippet: Following blocking, membranes were incubated overnight at 4°C with primary antibodies against β-Actin (1:2000; Proteintech, Chicago, IL, USA), C1QBP (1:1000; Abcam), TRAF2 (1:1000; Proteintech), and CCL2 (1:500; Proteintech).

Techniques: Knockdown, Expressing, Quantitative RT-PCR

Journal: International Dental Journal

Article Title: C1QBP Drives M2 Macrophage Polarization Via TRAF2-CCL2 to Promote Oral Squamous Cell Carcinoma Progression

doi: 10.1016/j.identj.2025.103938

Figure Lengend Snippet: C1QBP regulates macrophage polarization through the TRAF2-CCL2 signaling axis in OSCC.

Article Snippet: Following blocking, membranes were incubated overnight at 4°C with primary antibodies against β-Actin (1:2000; Proteintech, Chicago, IL, USA), C1QBP (1:1000; Abcam), TRAF2 (1:1000; Proteintech), and CCL2 (1:500; Proteintech).

Techniques:

Journal: bioRxiv

Article Title: Epstein-Barr Virus Latent Membrane Protein 1 targets cIAP1, cIAP2 and TRAF2 for Proteasomal Degradation to Activate the Non-canonical NF-κB Pathway

doi: 10.1101/2025.09.06.674652

Figure Lengend Snippet: (A) Schematic diagram of wildtype (WT), TES1 point mutant (TES1m), TES2 point mutant (TES2m), or TES1/2 double mutant (DM) LMP1. (B) Analysis of LMP1 effects on cIAP1, cIAP2, XIAP, and TRAFs expression in Daudi Burkitt cells. Immunoblot analysis of whole cell lysates (WCL) from Cas9+ Daudi Burkitt B-cells induced for WT, TES1m, TES2m, or DM LMP1 expression by addition of 250ng/mL doxycycline (Dox) for 24 hours. (C) Relative fold changes + standard deviation (SD) of GAPDH load control normalized cIAP1, cIAP2 and TRAF2 levels, based on densitometry from three replicates as in (B). Values in vehicle control treated WT LMP1 expressing cells were set to 1. (D) Analysis of LMP1 effects on cIAP1 and cIAP2 expression in newly infected primary B-cells. Immunoblot analysis of WCL from human peripheral blood B cells infected with EBV expressing WT, TES1m, TES2m or DM LMP1 at day 7 post-infection. DDX1 was used as a load control, since its protein levels do not change significantly following primary B cell infection by EBV[ , ]. (E) Relative fold changes + SD of GAPDH load control normalized cIAP1 and cIAP2 levels, based on densitometry from three replicates as in (D). Values in vehicle control treated WT LMP1 expressing cells were set to 1. Statistical significance was assessed by two-tailed unpaired Student’s t-test (C) or one-way ANOVA followed by Tukey’s multiple comparisons test (E). ns, not significant, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Blots (B and D) are representative of n=3 experiments.

Article Snippet: Antibodies against TRAF2 (Proteintech 26846-1-AP), TRAF2 (Invitrogen SD205-06), HA (Abcam ab9110), and p100/52 (EMD Millipore #05-361) were used at 1:1000.

Techniques: Mutagenesis, Expressing, Western Blot, Standard Deviation, Control, Infection, Two Tailed Test

Journal: bioRxiv

Article Title: Epstein-Barr Virus Latent Membrane Protein 1 targets cIAP1, cIAP2 and TRAF2 for Proteasomal Degradation to Activate the Non-canonical NF-κB Pathway

doi: 10.1101/2025.09.06.674652

Figure Lengend Snippet: (A) Analysis of cIAP1, cIAP2, and TRAF2 half-lives in isogenic MUTU Burkitt B-cells that differ only by EBV latency I vs III programs. Shown are representative immunoblot analyses of WCL from MUTU I vs MUTU III cells treated with 50 µg/mL cycloheximide (CHX) for the indicated number of hours. Blots are representative of n=3 experiments. (B) Relative fold changes + SD of GAPDH load control normalized cIAP1, cIAP2 and TRAF2 levels, based on densitometry from three replicates as in (A). Values in MUTU I and III cells at 0 hours of CHX chase were set to 1. Statistical significance was assessed by two-way ANOVA followed by Tukey’s multiple comparisons test (B). ns, not significant, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: Antibodies against TRAF2 (Proteintech 26846-1-AP), TRAF2 (Invitrogen SD205-06), HA (Abcam ab9110), and p100/52 (EMD Millipore #05-361) were used at 1:1000.

Techniques: Western Blot, Control

Journal: bioRxiv

Article Title: Epstein-Barr Virus Latent Membrane Protein 1 targets cIAP1, cIAP2 and TRAF2 for Proteasomal Degradation to Activate the Non-canonical NF-κB Pathway

doi: 10.1101/2025.09.06.674652

Figure Lengend Snippet: (A) Analysis of TRAF3 roles in LMP1 TES1-mediated cIAP1 and cIAP2 depletion. Immunoblot analysis of WCL from Cas9+ Daudi cells that expressed control vs TRAF3 targeting single guide RNA (sgRNA) and that were induced for LMP1 expression by Dox (250 ng/mL) for 24 hours. (B) Relative fold changes + SD of GAPDH load control normalized cIAP1, cIAP2, based on densitometry from three immunoblot replicates as shown in (A). Values in Daudi cells with control sgRNA and uninduced for WT LMP1 expression were set to 1. (C) Analysis of cIAP1, cIAP2, and TRAF2 half-lives in control vs TRAF3 knockout (KO) GM12878 LCLs. Immunoblot analysis of WCL from Cas9+ GM12878 LCLs that expressed control versus TRAF3 targeting sgRNA and that were treated with 50µg/mL CHX for the indicated hours (hrs). (D) Relative fold changes + SD of GAPDH load control normalized cIAP1, cIAP2 and TRAF2 levels, based on densitometry from three replicates as in (C). Values in GM12878 cells with control sgRNA expression at 0 hours of CHX chase were set to 1. Statistical significance was assessed by two-tailed unpaired Student’s t-test (B) or by two-way ANOVA followed by Tukey’s multiple comparisons test (D). ns, not significant, *p<0.05, **p<0.01, ****p<0.0001. Blots are representative of n=3 experiments.

Article Snippet: Antibodies against TRAF2 (Proteintech 26846-1-AP), TRAF2 (Invitrogen SD205-06), HA (Abcam ab9110), and p100/52 (EMD Millipore #05-361) were used at 1:1000.

Techniques: Western Blot, Control, Expressing, Knock-Out, Two Tailed Test

Journal: bioRxiv

Article Title: Epstein-Barr Virus Latent Membrane Protein 1 targets cIAP1, cIAP2 and TRAF2 for Proteasomal Degradation to Activate the Non-canonical NF-κB Pathway

doi: 10.1101/2025.09.06.674652

Figure Lengend Snippet: Under unstimulated conditions, the TRAF-cIAP complex targets the kinase NIK for proteasomal degradation, thereby preventing non-canonical NF-κB signaling. LMP1 TES1 triggers TRAF2, cIAP1 and cIAP2 for degradation, which initiates downstream signaling and p100:p52 processing. By comparison, CD40L/CD40 signaling induces degradation of TRAF3 to drive downstream non-canonical NF-kB pathway activation.

Article Snippet: Antibodies against TRAF2 (Proteintech 26846-1-AP), TRAF2 (Invitrogen SD205-06), HA (Abcam ab9110), and p100/52 (EMD Millipore #05-361) were used at 1:1000.

Techniques: Comparison, Activation Assay