Journal: Nature Communications

Article Title: Endothelin 3/EDNRB signaling induces thermogenic differentiation of white adipose tissue

doi: 10.1038/s41467-024-51579-0

Figure Lengend Snippet: a – d Human white preadipocytes were pretreated with vehicle (Veh) or 100 nM EDN3 and with or without inhibitors for 3 days, followed by a 12-day adipogenesis. a Schematic of pretreatment experiments. UCP1 mRNA ( b ) and protein ( c ) levels in mature adipocytes (EV and EV and EDNRB OE). d UCP1 mRNA levels in mature adipocytes (EV and EDNRB KO). e Cytosolic calcium levels in human white preadipocytes after 100 nM EDN3 treatment. f , g Intracellular cAMP levels ( f ) and oxygen consumption rate (OCR; g ) in human white preadipocytes after 6 hours of Veh or EDN3 treatment. Maximal OCR was quantified in the right panel. h – j EV and EDNRB OE preadipocytes were pretreated with Veh or EDN3 combined with or without melittin (ME) or YM254890 (YM) for 3 days, followed by a 12-day adipogenesis. UCP1 mRNA ( h ) and protein ( i ) levels in mature adipocytes pretreated with ME. j UCP1 mRNA levels in mature adipocytes pretreated with YM. k The protein levels in preadipocytes after 6 hr of Veh or EDN3 treatment. The quantification of protein bands was in the right panels. l The EPAC1 protein in EDNRB OE preadipocytes after 48 hr of EPAC1 siRNA treatment. m EDNRB OE preadipocytes with or without EPAC1 knockdown were pretreated with Veh or EDN3 for 3 days, followed by a 12-day adipogenesis. Protein levels in mature adipocytes. n EV or EDNRB OE preadipocytes were pretreated with Veh, 1 μM or 10 μM of PD98059 (PD-1 or PD-10) for 3 days and were differentiated to mature adipocytes. UCP1 and DIO2 mRNA levels in human white adipocytes. n = 3 biological replicates/group in ( b – e ), ( i ), ( k – n ); n = 4 biological replicates/group in ( f – h ), ( j ). Data are presented as mean ± SEM. P values were determined using unpaired two-tailed t tests: ( l ); one-way ANOVA with Tukey’s multiple-comparison test: ( b – d ), ( f – k ), ( m ), ( n ); two-way ANOVA with Bonferroni’s multiple-comparison test: ( e ). Source data are provided as a Source Data file. Panel ( a ) was created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license.

Article Snippet: Primary antibodies for EDNRB (NBP130599, Novus Biologicals), UCP1 (ab23841, abcam), phospho-ERK1/2 (Thr202/Tyr204) (#4370, Cell Signaling), total ERK1/2 (#4695, Cell Signaling), phospho-CREB (Ser133) (#9198, Cell Signaling), total CREB (#9197, Cell Signaling), Phospho-PKA Substrate (#9624, Cell Signaling), PPARγ (GTX32803, GeneTex), FABP4/AP2 (GTX636995, GeneTex), EPAC1 (#4155, Cell Signaling), TUBULIN (#2146, Cell Signaling), VINCULIN (sc-25336, Santa Cruz) or ACTIN (MAB1501, Millipore) were applied in blocking buffer over night at 4°C.

Techniques: Knockdown, Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: Endothelin 3/EDNRB signaling induces thermogenic differentiation of white adipose tissue

doi: 10.1038/s41467-024-51579-0

Figure Lengend Snippet: a , b Male mice were intraperitoneally injected by tamoxifen 5 times within 7 days to induce EDNRB KO in PDGFRα + preadipocytes (iKO) and were then housed at 5 °C for 7 days. a The mRNA levels of thermogenic genes in scWAT. CTL: n = 5, iKO: n = 6. b The protein levels of UCP1 in scWAT. The quantification of protein bands in right panel. n = 4 mice/group. c , d Control and iKO mice were housed at thermoneutral temperature (30 °C, TN) or cold (5 °C) for 7 days. Images of immunohistochemical stain for UCP1 proteins (UCP1 IHC; c ) and HE stain ( d ) in scWAT. High magnification images were in the right panels. Scale bar=100 μm. Two independent experiments were repeated with similar results, as shown in Supplementary Fig. a and . e The phosphorylated ERK and total ERK protein in scWAT of mice after 7 days of cold exposure. The quantification of protein bands in right panel. n = 3 mice/group. f , g PDGFRα + cell lineage-tracing control and EDNRB iKO mice were housed at thermoneutral temperature (30 °C, TN) or cold (5°C) for 7 days. f The immunofluorescent images from the section of scWAT. Scale bar=100 μm. g Quantification of GFP signal in each section of scWAT. CTL at TN: n = 6, iKO at TN: n = 6, CTL at Cold: n = 6, iKO at Cold: n = 7, 4 slides/mouse. h The mRNA levels of Ednrb, Pdgfrα and Ebf2 in SVF of scWAT from mice housed at TN or cold for 7 days. CTL at TN: n = 3, iKO at TN: n = 3, CTL at Cold: n = 3, iKO at Cold: n = 4. Data are presented as mean ± SEM. P values were determined using unpaired two-tailed t tests: ( a) , ( b ), ( e ); one-way ANOVA with Tukey’s multiple-comparison test: ( g ), ( h ). Source data are provided as a Source Data file.

Article Snippet: Primary antibodies for EDNRB (NBP130599, Novus Biologicals), UCP1 (ab23841, abcam), phospho-ERK1/2 (Thr202/Tyr204) (#4370, Cell Signaling), total ERK1/2 (#4695, Cell Signaling), phospho-CREB (Ser133) (#9198, Cell Signaling), total CREB (#9197, Cell Signaling), Phospho-PKA Substrate (#9624, Cell Signaling), PPARγ (GTX32803, GeneTex), FABP4/AP2 (GTX636995, GeneTex), EPAC1 (#4155, Cell Signaling), TUBULIN (#2146, Cell Signaling), VINCULIN (sc-25336, Santa Cruz) or ACTIN (MAB1501, Millipore) were applied in blocking buffer over night at 4°C.

Techniques: Injection, Control, Immunohistochemical staining, Staining, H&E Stain, Two Tailed Test, Comparison

Journal: Translational Oncology

Article Title: Diagnostic and prognostic value of EphB2 in nasopharyngeal carcinoma

doi: 10.1016/j.tranon.2025.102641

Figure Lengend Snippet: Comparison of serum EphB2 levels between NPC patients and the control group. Data are presented as mean ± SD.

Article Snippet: The membranes were then incubated with a primary antibody against EphB2 (BOSTER, #A01507–1) overnight at 4 °C, followed by incubation with an HRP-conjugated secondary antibody.

Techniques: Comparison, Control

Journal: Translational Oncology

Article Title: Diagnostic and prognostic value of EphB2 in nasopharyngeal carcinoma

doi: 10.1016/j.tranon.2025.102641

Figure Lengend Snippet: EphB2 protein expression in nasopharyngeal tissues. (A) Representative western blot images of EphB2 expression in control tissues (nasopharyngeal mucosal chronic inflammation) and NPC tissues stratified by clinical stage (I–II vs. III–IV), EBV status, and recurrence status ( n = 3 per subgroup). Approximate molecular weights: EphB2, 117 kDa; GAPDH, 36 kDa. (B) Quantitative analysis of EphB2 expression normalized to GAPDH. Data represent mean ± SD from six independent experiments. ** P < 0.01 by one-way ANOVA with Tukey’s post-hoc test.

Article Snippet: The membranes were then incubated with a primary antibody against EphB2 (BOSTER, #A01507–1) overnight at 4 °C, followed by incubation with an HRP-conjugated secondary antibody.

Techniques: Expressing, Western Blot, Control

Journal: Translational Oncology

Article Title: Diagnostic and prognostic value of EphB2 in nasopharyngeal carcinoma

doi: 10.1016/j.tranon.2025.102641

Figure Lengend Snippet: Immunofluorescence analysis of EphB2 expression in nasopharyngeal tissues. A: Nasopharyngeal mucosal chronic inflammation tissues; B: EBV(-) NPC tissues; C: EBV(+) NPC tissues. EphB2 staining is shown in red; nuclei are counterstained with DAPI (blue). Original magnification: 400×.

Article Snippet: The membranes were then incubated with a primary antibody against EphB2 (BOSTER, #A01507–1) overnight at 4 °C, followed by incubation with an HRP-conjugated secondary antibody.

Techniques: Immunofluorescence, Expressing, Staining

Journal: Translational Oncology

Article Title: Diagnostic and prognostic value of EphB2 in nasopharyngeal carcinoma

doi: 10.1016/j.tranon.2025.102641

Figure Lengend Snippet: Disease-free survival analysis based on recurrence (A), EBV infection status (B), and serum EphB2 expression (C). In (C), patients were divided into EphB2-Low and EphB2-High groups using the median serum concentration (11.161 ng/mL) as the cutoff. Vertical ticks indicate censored data.

Article Snippet: The membranes were then incubated with a primary antibody against EphB2 (BOSTER, #A01507–1) overnight at 4 °C, followed by incubation with an HRP-conjugated secondary antibody.

Techniques: Infection, Expressing, Concentration Assay

Journal: Translational Oncology

Article Title: Diagnostic and prognostic value of EphB2 in nasopharyngeal carcinoma

doi: 10.1016/j.tranon.2025.102641

Figure Lengend Snippet: ROC curves evaluating the diagnostic performance of serum EphB2, EBV infection status, and their combination in discriminating NPC patients from controls.

Article Snippet: The membranes were then incubated with a primary antibody against EphB2 (BOSTER, #A01507–1) overnight at 4 °C, followed by incubation with an HRP-conjugated secondary antibody.

Techniques: Diagnostic Assay, Infection