Journal: Biology of Sex Differences

Article Title: Sex differences in disease severity and immune responses in murine and human inflammatory arthritis

doi: 10.1186/s13293-026-00840-w

Figure Lengend Snippet: Male CIA mice exhibit higher disease severity compared to females. Male and female CIA and saline control mice were monitored for disease severity and assigned clinical scores from day 21 after the first CII challenge until the end of experiment (day 29). (A) Line graphs representing the mean clinical scores of mice starting from day 1 to day 29. On day 29 after the first CII challenge, mice were euthanized by cardiac puncture under anesthesia for blood and serum collection and storage at −80 °C. Serum concentrations of (B) anti-mouse collagen type II antibodies (left panel) and anti-bovine collagen type II antibodies (right panel) were analyzed by ELISA. N = 5 mice per group. One of two independent experiments. Simple linear regression analysis was performed to determine the statistical difference between the lines. One-way analysis of variance (ANOVA) followed by Tukey’s multiple comparison post hoc test was used to determine the statistical significance between the groups. (*/ # p ≤ 0.05, ** p ≤ 0.005 and ***/ ### p ≤ 0.0005). In Fig. 1A, # significance of comparisons between CIA and saline control mice; *significance of comparisons between sexes of CIA mice

Article Snippet: Serum levels of mouse anti-collagen antibodies (autoantibodies) and bovine anti-collagen antibodies (antibodies to the immunizing antigen) were determined by ELISA using a Mouse Anti-mouse Type II Collagen IgG Antibody Assay Kit and Mouse Anti-Bovine Type II Collagen IgG Antibody Assay Kit, respectively, according to the manufacture’s protocol (Chondrex Inc. WA, USA).

Techniques: Saline, Control, Enzyme-linked Immunosorbent Assay, 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 – 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: Materials today. Bio

Article Title: Precise delivery of doxorubicin and imiquimod through pH-responsive tumor microenvironment-active targeting micelles for chemo- and immunotherapy.

doi: 10.1016/j.mtbio.2022.100482

Figure Lengend Snippet: Fig. 10. Immunostaining of tumor tissues in tumor-bearing mice after treatments. (A) Immunohistochemistry images of tumor sections stained with CD3, CD8, and TNF-α antibodies. The scale bar is 100 μm. (B) Immunofluorescence images of tumor tissues after treatments for 12 days. The scale bar is 50 μm. Blue fluorescence represents the cell nucleus stained with DAPI. Green fluorescence represents the iNOS stained with the iNOS antibody conjugated with FITC.

Article Snippet: After 30 min, the tissue slice was stained with diluted iNOS antibody (Miltenyi Biotec, catalog: 130-116-357) at 4 C overnight.

Techniques: Immunostaining, Immunohistochemistry, Staining