Journal: Poultry Science

Article Title: METTL14-mediated RNA m 6 A modifications of TET1 and TET2 in duck myoblast differentiation

doi: 10.1016/j.psj.2026.106435

Figure Lengend Snippet: Comparison of RNA m 6 A modification levels in duck leg muscle. (A) DPMs during the proliferation phase; (B) DPMs on day 2 of differentiation (myotubes); (C) Immunofluorescence for Desmin in DPMs during proliferation; (D) Immunofluorescence for MYHC in DPMs on day 4 of differentiation. Dot blot and quantitative analysis of m 6 A in total RNA from duck embryos at E10 and E20 are shown in panels (E) and (F); panels (G) and (H) display quantitative and dot blot analyses of m 6 A in total RNA from DPMs in the GM and DM phases. Methylene blue (MB) staining served as a control for RNA loading. E10 and E20 indicate leg muscle samples from duck embryos at 10 and 20 days of incubation, respectively. GM and DM represent undifferentiated DPMs and DPMs at 48 hours post-differentiation, respectively. * indicates P < 0.05, ** indicates P < 0.01.

Article Snippet: The membrane was blocked in 5% nonfat milk in PBS for 2 hours, followed by overnight incubation at 4°C with primary antibodies against MYHC (DSHB, Iowa, IA, USA; 0.3 μg/mL) and GAPDH (Proteintech, Wuhan, China; 1:50000).

Techniques: Comparison, Modification, Immunofluorescence, Dot Blot, Staining, Control, Incubation

Journal: Biochemistry and Biophysics Reports

Article Title: Accumulation of prosaposin and progranulin around the subfornical organ induces polydipsia in SAP-D-deficient mice

doi: 10.1016/j.bbrep.2025.102388

Figure Lengend Snippet: Increased protein levels of PSAP and PGRN within the SFO of SAP-D −/− mice a – b) , Coronal brain section at 0.7–0.8 mm posterior to bregma containing the SFO. a: DAPI staining. a-2: Enlarged view of the white square in a-1. SFO: subfornical organ, 3V: third ventricle. b; Double immunofluorescent staining of PSAP (red) and PGRN (green) in 10-month-old female WT and SAP-D −/− mice. DAPI (blue) staining showed the nuclei ( a and b ). Scale bar, 500 μm ( a and b ). White arrowheads indicate the SFO regions ( b ). c) Cerebral region from 3-, 6-, and 10-month-old male and female mice containing the SFO (0.7–0.8 mm posterior to ∗bregma) used for protein extraction and Western blot ( d – h ) using anti- PSAP, PGRN, and GAPDH antibodies. Quantification normalized to GAPDH expression and represented as the mean ± SD of three mice for each group. and indicate the individual values in each group ( e , f , h ). d – f) PSAP and PGRN protein levels in the SFO were remarkably increased. Their quantification by densitometric analysis is represented in e for male and f for female, respectively. e ) For PSAP/GAPDH, two-way ANOVA revealed a significant main effects of genotype (F(1,24) = 547.7, p < 0.0001, ηp 2 = 0.48, 95 % CI [−74.51, −62.43]), with no effect of age ( p = 0.46) or genotype × age interaction ( p = 0.47). Post-hoc Tukey's tests showed that SAP-D −/− differed from WT at 3 M ( p < 0.0001, Cohen's d = 9.99, 95 % CI [−86.30, −54.96]), 6 M ( p < 0.0001, Cohen's d = 7.24, 95 % CI [−79.04, −47.70]), and 10 M ( p < 0.0001, Cohen's d = 8.77, 95 % CI [−87.09, −55.75]). For PGRN/GAPDH, two-way ANOVA revealed a significant main effect of genotype (F(1,24) = 354.2, p < 0.0001, ηp 2 = 0.48, 95 % CI [−46.39, −37.22]), with no effect of age ( p = 0.73) or genotype × age interaction ( p = 0.76). Post-hoc Tukey's tests showed that SAP-D −/− differed from WT at 3 M ( p < 0.0001, Cohen's d = 7.87, 95 % CI [−52.71, −28.91]), 6 M ( p < 0.0001, Cohen's d = 5.35, 95 % CI [−52.82, −29.02]), and 10 M ( p < 0.0001, Cohen's d = 8.77, 95 % CI [−52.32, −28.52]). f ) For PSAP/GAPDH, two-way ANOVA revealed a significant main effect of genotype (F(1,24) = 611.1, p < 0.0001, ηp 2 = 0.48, 95 % CI [−74.42, −62.95]), with no effect of age ( p = 0.07) or genotype × age interaction (p = 0.07). Post-hoc Tukey's tests showed that SAP-D −/− differed from WT at 3 M ( p < 0.0001, Cohen's d = 9.99, 95 % CI [−86.30, −54.96]), 6 M ( p < 0.0001, Cohen's d = 7.24, 95 % CI [−79.04, −47.70]), and 10 M ( p < 0.0001, Cohen's d = 8.77, 95 % CI [−87.09, −55.75]). For PGRN/GAPDH, two-way ANOVA revealed a significant main effect of genotype (F(1,24) = 273.5, p < 0.0001, ηp 2 = 0.47, 95 % CI [−63.96, −49.76]), with no effect of age ( p = 0.71) or genotype × age interaction (p = 0.70). Post-hoc Tukey's tests showed that SAP-D −/− differed from WT at 3 M ( p < 0.0001, Cohen's d = 7.31, 95 % CI [−73.43, −36.60]), 6 M ( p < 0.0001, Cohen's d = 5.71, 95 % CI [−73.04, −36.22]), and 10 M ( p < 0.0001, Cohen's d = 5.35, 95 % CI [−73.08, −36.26]). g – h ) Comparison of PSAP and PGRN protein expression in the SFO, whole cerebrum, and cerebellum. The quantitative analysis is shown in h . h ) For PSAP/GAPDH, one-way ANOVA revealed a significant effect in SAP-D −/− mice (F(2,6) = 30.06, p = 0.0007, η 2 = 0.90), but not in WT mice ( p = 0.3461). Tukey's post hoc tests showed significant differences for SFO versus cerebellum ( p = 0.0010, Cohen's d = 5.26, 95 % CI [46.80, 119.1]) as well as and cerebrum versus cerebellum ( p = 0.0018, Cohen's d = 9.34, 95 % CI [38.39, 110.6]). There was no significant difference for SFO versus cerebrum ( p = 0.76, Cohen's d = 0.47, 95 % CI [−27.71, 44.54]). For PGRN/GAPDH, one-way ANOVA revealed a significant effect in SAP-D −/− mice (F(2,6) = 54.42, p = 0.0001, η 2 = 0.94), but not in WT mice ( p = 0.6327). Tukey's post hoc tests showed significant differences for SFO versus cerebrum ( p = 0.0005, Cohen's d = 5.43, 95 % CI [26.19, 57.84]) as well as and SFO versus cerebellum ( p = 0.0005, Cohen's d = 5.43, 95 % CI [26.19, 57.84]), but not for cerebrum versus cerebellum ( p = 0.3274, Cohen's d = 6.06, 95 % CI [−7.72, 23.92]). ns: no significant difference. ∗∗∗∗ p < 0.0001. ∗∗∗ p < 0.001. ∗∗ p < 0.01.

Article Snippet: The primary antibodies used were a rabbit antibody against PSAP (dilution 1:100, 10801-1-AP, Proteintech Group Inc., IL, USA), a sheep antibody against PGRN (dilution 1:100, AF 2557, R&D Systems Inc., MN, USA), a guinea pig antibody against c-Fos (dilution 1:500, 226308, Synaptic Systems GmbH, Göttingen, Germany), a rat antibody against CD68-FITC conjugated (dilution 1:500, MCA1957FA, BIO-RAD Laboratories Inc., CA, USA), and a rat antibody against LAMP1 (dilution 1:100, ab25245, Abcam, Cambridge, UK).

Techniques: Staining, Protein Extraction, Western Blot, Expressing, Comparison

Journal: Biochemistry and Biophysics Reports

Article Title: Accumulation of prosaposin and progranulin around the subfornical organ induces polydipsia in SAP-D-deficient mice

doi: 10.1016/j.bbrep.2025.102388

Figure Lengend Snippet: Increased PSAP and PGRN immunostaining in the SFO and its surrounding tissues in SAP-D −/− mice a) Double immunofluorescent staining of PSAP (red) and PGRN (green) around the SFO in 10-month-old female WT and SAP-D −/− mice. The white dotted lines enclose the SFO. b ) enlarged white ⅰ-iv squares in a, as indicated. White arrowheads indicate co-staining with anti-PGRN and PSAP antibodies. Open arrowheads indicate staining with PGRN alone. Nuclei are labeled by DAPI (blue) staining. All scale bars, 20 μm.

Article Snippet: The primary antibodies used were a rabbit antibody against PSAP (dilution 1:100, 10801-1-AP, Proteintech Group Inc., IL, USA), a sheep antibody against PGRN (dilution 1:100, AF 2557, R&D Systems Inc., MN, USA), a guinea pig antibody against c-Fos (dilution 1:500, 226308, Synaptic Systems GmbH, Göttingen, Germany), a rat antibody against CD68-FITC conjugated (dilution 1:500, MCA1957FA, BIO-RAD Laboratories Inc., CA, USA), and a rat antibody against LAMP1 (dilution 1:100, ab25245, Abcam, Cambridge, UK).

Techniques: Immunostaining, Staining, Labeling

Journal: Biochemistry and Biophysics Reports

Article Title: Accumulation of prosaposin and progranulin around the subfornical organ induces polydipsia in SAP-D-deficient mice

doi: 10.1016/j.bbrep.2025.102388

Figure Lengend Snippet: Lysosomal localization of PSAP and PGRN expression around the SFO a) Immunofluorescence staining of LAMP1 (green) around the SFO in 10-month-old female WT and SAP-D −/− mice. Nuclei are labeled by DAPI (blue) staining. b) Quantification of LAMP1-stained areas in ( a ) relative to WT (%). The results of Student's t-tests for each panel are as follows: left panel, p = 0.0059, Cohen's d = 2.35 (95 % CI: 15.89, 67.66); and right panel: p = 0.0061, Cohen's d = 2.33 (95 % CI: 51.06, 220.89). Data are shown as the mean ± SD (n = 5). c) Triple immunofluorescent staining of PSAP (red), PGRN (green), and LAMP1 (cyan) around the SFO of 10-month-old female SAP-D −/− mice. Enlarged images indicated by the white squares (ⅰ-iv) were shown in c. d) Localization rate of PGRN or PSAP to LAMP1 in the SFO. The results of Student's t-tests for each panel are as follows: left panel, p = 0.0008, Cohen's d = 2.24 (95 % CI: -40.90, -13.73); and right panel, p = 0.2341, Cohen's d = 0.64 (95 % CI: 26.80, 7.17). Data are shown as the mean ± SD (n = 6 for WT-SFO and n = 8 for SAP-D −/− -SFO). e) Same experiment as in (c) on microglia/macrophage co-expressing PSAP and PGRN, or PGRN only, in the boundary and fornix regions of SAP-D −/− mice. Single image for each antibody was shown in white, while double or triple merged images were presented with red (PSAP or PGRN), green (LAMP1 or PGRN), or cyan (LAMP1) as indicated. and indicate the individual values in each group (b and d). ns: no significant difference. ∗∗∗ p < 0.001. ∗∗ p < 0.01. All scale bars, 10 μm.

Article Snippet: The primary antibodies used were a rabbit antibody against PSAP (dilution 1:100, 10801-1-AP, Proteintech Group Inc., IL, USA), a sheep antibody against PGRN (dilution 1:100, AF 2557, R&D Systems Inc., MN, USA), a guinea pig antibody against c-Fos (dilution 1:500, 226308, Synaptic Systems GmbH, Göttingen, Germany), a rat antibody against CD68-FITC conjugated (dilution 1:500, MCA1957FA, BIO-RAD Laboratories Inc., CA, USA), and a rat antibody against LAMP1 (dilution 1:100, ab25245, Abcam, Cambridge, UK).

Techniques: Expressing, Immunofluorescence, Staining, Labeling

Journal: International Journal of Molecular Sciences

Article Title: Hedyotis diffusa Suppresses Colitis-Associated Colorectal Cancer via Inhibition of the IL-17A-IL-17RA Axis and NF-κB Signaling

doi: 10.3390/ijms27041745

Figure Lengend Snippet: Molecular docking of FA with the IL-17A/IL-17RA complex. The overall docking model shows FA (red) binding at the interface between IL-17A (blue) and IL-17RA (green). The enlarged panel shows a close-up view of the interaction between FA and IL-17A chain B, where hydrogen bonds are indicated by yellow dashed lines.

Article Snippet: Mouse monoclonal antibody against p65 (MAB3026), rabbit polyclonal antibody against IL-17A (bs-1183R), and rabbit monoclonal antibody against CD11b (ab133357) were obtained from Millipore (Temecula, CA, USA), Bioss Antibodies (Woburn, MA, USA), and abcam (Cambridge, UK), respectively.

Techniques: Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Hedyotis diffusa Suppresses Colitis-Associated Colorectal Cancer via Inhibition of the IL-17A-IL-17RA Axis and NF-κB Signaling

doi: 10.3390/ijms27041745

Figure Lengend Snippet: Effects of HD and FA on NF-κB activation, IL-17A production, and granulocyte infiltration in rectal tissues of AOM/DSS-induced CRC mice. Mice were treated with HD (200 mg/kg) or FA (100 mg/kg) for 91 days. Rectal sections were stained with antibody against p65, IL-17A, or CD11b (200× magnification). Scale bar = 100 µm. Representative images are shown. Quantitative analysis of the stained area or cells (%) is displayed below the image. Data are expressed as mean ± SD (n = 8 mice per group). ### p < 0.001 vs. mock group. ** p < 0.01, *** p < 0.001 vs. CRC group.

Article Snippet: Mouse monoclonal antibody against p65 (MAB3026), rabbit polyclonal antibody against IL-17A (bs-1183R), and rabbit monoclonal antibody against CD11b (ab133357) were obtained from Millipore (Temecula, CA, USA), Bioss Antibodies (Woburn, MA, USA), and abcam (Cambridge, UK), respectively.

Techniques: Activation Assay, Staining

Journal: bioRxiv

Article Title: Distinct activation thresholds of unmyelinated C-fiber afferents by dorsal root ganglion and peripheral nerve stimulation

doi: 10.64898/2026.02.02.703367

Figure Lengend Snippet: Analyses of two prior single-cell RNA-seq studies of mouse DRG neurons reveal widespread expression of NaV1.6 across both A- and C-fiber afferents , . (A) Violin plots showing NaV1.6 (Scn8a) mRNA expression levels across distinct DRG neuronal populations. NF, neurofilament; NP, non-peptidergic; PEP, peptidergic; TH, tyrosine hydroxylase. (B) Violin plot showing NaV1.6 mRNA expression in colonic DRG neurons. RPM, reads per million; TPM, transcripts per million.

Article Snippet: Tissue sections were incubated with rabbit antibodies against NaV1.6 (1:1000, Alomone) and co-stained with rat antibodies against GFP (1:1000, MBL International Corp) for 24 h at 4 °C, and then incubated with a mixture of secondary antibodies Alexa Fluor® 594-conjugated anti-rabbit IgG (1:200, Abcam) and Alexa Fluor® 488-conjugated anti-rat IgG (1:200, Abcam) for 2 h at room temperature.

Techniques: Single Cell, RNA Sequencing, Expressing

Journal: bioRxiv

Article Title: Distinct activation thresholds of unmyelinated C-fiber afferents by dorsal root ganglion and peripheral nerve stimulation

doi: 10.64898/2026.02.02.703367

Figure Lengend Snippet: Both large- and small-diameter DRG neurons show clustering of NaV1.6 in stem axons near the somata. (A) Representative DRG sections co-stained for NaV1.6 (red) and afferents (green) showing NaV1.6 clusters (indicated by white arrow heads) in the stem axon of both large- and small-diameter neurons. Scale bar: 20 μm. (B) Distance between the somata and the first NaV1.6 clustering in the stem axon. There was no significant difference between the large- and small-diameter groups (t-test, p = 0.58).

Article Snippet: Tissue sections were incubated with rabbit antibodies against NaV1.6 (1:1000, Alomone) and co-stained with rat antibodies against GFP (1:1000, MBL International Corp) for 24 h at 4 °C, and then incubated with a mixture of secondary antibodies Alexa Fluor® 594-conjugated anti-rabbit IgG (1:200, Abcam) and Alexa Fluor® 488-conjugated anti-rat IgG (1:200, Abcam) for 2 h at room temperature.

Techniques: Staining

Journal: bioRxiv

Article Title: Distinct activation thresholds of unmyelinated C-fiber afferents by dorsal root ganglion and peripheral nerve stimulation

doi: 10.64898/2026.02.02.703367

Figure Lengend Snippet: NaV1.6 clustering is present in A-fiber axons but absent in C-fiber axons of the sciatic nerve. (A) Representative immunostaining of NaV1.6 in sciatic nerve sections with sparsely labeled afferents following intrathecal AAV9 injection. Arrowheads indicate the presence of NaV1.6 clusters in Aβ- (Φ > 4.5 μm) and Aδ-fiber axons (1.5 μm < Φ < 4.5 μm), but not in C-fiber axons (Φ < 1.5 μm). (B) NaV1.6 cluster density (clusters per 100 μm) is significantly higher in Aβ-fiber axons than in Aδ-fiber axons (mean ± SEM; t test, p = 0.003). Scale bar: 10 μm.

Article Snippet: Tissue sections were incubated with rabbit antibodies against NaV1.6 (1:1000, Alomone) and co-stained with rat antibodies against GFP (1:1000, MBL International Corp) for 24 h at 4 °C, and then incubated with a mixture of secondary antibodies Alexa Fluor® 594-conjugated anti-rabbit IgG (1:200, Abcam) and Alexa Fluor® 488-conjugated anti-rat IgG (1:200, Abcam) for 2 h at room temperature.

Techniques: Immunostaining, Labeling, Injection

Journal: bioRxiv

Article Title: Distinct activation thresholds of unmyelinated C-fiber afferents by dorsal root ganglion and peripheral nerve stimulation

doi: 10.64898/2026.02.02.703367

Figure Lengend Snippet: NaV1.6 clustering in afferent nerve endings in the bladder. (A-B) Immunostaining of bladder tissue sections from VGLUT2/tdTomato mice showed the presence of NaV1.6 clustering, in some bladder afferent axons (putative nerve terminals) as indicated by white arow heads. (C–D) Non-terminal axons of bladder afferents lack detectable NaV1.6 staining. Scale bar: 20 μm.

Article Snippet: Tissue sections were incubated with rabbit antibodies against NaV1.6 (1:1000, Alomone) and co-stained with rat antibodies against GFP (1:1000, MBL International Corp) for 24 h at 4 °C, and then incubated with a mixture of secondary antibodies Alexa Fluor® 594-conjugated anti-rabbit IgG (1:200, Abcam) and Alexa Fluor® 488-conjugated anti-rat IgG (1:200, Abcam) for 2 h at room temperature.

Techniques: Immunostaining, Staining

Journal: bioRxiv

Article Title: Distinct activation thresholds of unmyelinated C-fiber afferents by dorsal root ganglion and peripheral nerve stimulation

doi: 10.64898/2026.02.02.703367

Figure Lengend Snippet: NEURON simulation of ePNS and DRG stimulation of a C-fiber afferent. (A) Action potentials evoked by DRG stimulation initiates close to the soma and propagates bi-directionally to both peripheral and central axons. (B) Action potentials evoked by ePNS of the peripheral axon propagates to soma and central axon. (C) The clustering of NaV1.6 in the stem axon strongly decreased the threshold of DRG stimulation to evoke APs. In contrast, comparable increases in NaV1.8 conductance within the clustering zone had no effect on the threshold, whereas increases in NaV1.7 conductance produced only a modest reduction. (D) Spatial plot along the soma and stem axon to reveal the site of spike initiation at the NaV1.6 clustering zone. Five spatial membrane voltage traces have 0.05 msec intervals (from 1 to 5).

Article Snippet: Tissue sections were incubated with rabbit antibodies against NaV1.6 (1:1000, Alomone) and co-stained with rat antibodies against GFP (1:1000, MBL International Corp) for 24 h at 4 °C, and then incubated with a mixture of secondary antibodies Alexa Fluor® 594-conjugated anti-rabbit IgG (1:200, Abcam) and Alexa Fluor® 488-conjugated anti-rat IgG (1:200, Abcam) for 2 h at room temperature.

Techniques: Produced, Membrane