mouse anti mhc  (Developmental Studies Hybridoma Bank)

Journal: bioRxiv

Article Title: Wnt/β-catenin signaling promotes zebrafish osteoblast dedifferentiation by wnt10a -mediated inhibition of NF-κB

doi: 10.64898/2025.12.29.696582

Figure Lengend Snippet: A) RNASeq (left) and RT-qPCR (right) reveal upregulation of the NF-κB signaling target genes nfkbiaa and nfkbiab in osteoblasts sorted from bglap:GFP fish heterozygous for the wnt10a mutation relative to their wild-type siblings at 1 dpa. nE (qPCR biological replicates) = 3, nA = 10 per replicate. ΔΔCt values are normalized to the mean of the wildtype at 1 dpa. Error bars, Mean ± SEM. Two tailed Student’s t-test. B) Osteoblast dedifferentiation, as measured by bglap downregulation in segment -1 revealed by HCR in situ hybridization, is inhibited in fish treated with the Wnt inhibitor IWR-1, while IP injection of the NF-κB inhibitor Bay-11 slightly but significantly enhances dedifferentiation. Treatment with both inhibitors yields results similar to those of Bay-11 alone. nE = 3 (except for 2 for IWR-1), nA = 18 total per group (12 for IWR-1), nR = 33 (DMSO), 24 (IWR-1), 37 (Bay-11), 33 (both). C) Overexpression of wnt10a using hs:wnt10a fish is sufficient to cause downregulation of bglapl detected by HCR in situ hybridization in non-injured fins, relative to heat-shocked wild-type fish. HCR signals were quantified in a bony segment (“B”) that is located at the same proximal-distal position as “segment -1” in amputated fins. nE = 2, nA = 12 total per group, nR = 22 total per group. Dashed line, joints. Scale bar, 100 µm. D) Immunofluorescence on cryosections of hs:wnt10a transgenic hearts reveals increased embryonic myosin heavy chain (embMHC) expression in Myl7+ cardiomyocytes at the wound border at 7 days post injury (dpi). Plots show the ventricular area covered by anti-embMHC staining relative to the 150 µm wound border zone area occupied by Myl7+ myocardium. nE = 2, nA = 13 wild-type, 11 hs:wnt10a . Scale bar, 100 µm. (B, C, D) Error bars, mean ± 95% CI. Two tailed Student’s t-test.

Article Snippet: Primary antibodies against embMHC Mouse monoclonal (MYH7 DSHB, N2.261, RRID:AB_531790), and Myl7 Rabbit polyclonal (GeneTex, GTX128346, RRID:AB_2885759) were diluted in PEMTx/normal goad serum and applied overnight at 4 °C.

Techniques: Quantitative RT-PCR, Mutagenesis, Two Tailed Test, In Situ Hybridization, Injection, Over Expression, Immunofluorescence, Transgenic Assay, Expressing, Staining

Journal: iScience

Article Title: Angiogenesis-independent VEGF signaling enhances exercise capacity by increasing fat oxidation in mice fed sulfur amino acid-restricted diets

doi: 10.1016/j.isci.2025.114148

Figure Lengend Snippet: Inhibition of VEGFR signaling blocks increased endurance exercise capacity by SAAR (A) Fold changes of transcripts associated with Vegf signaling using transcriptomic dataset presented in in muscles of male mice ( n = 6) after sulfur amino acid restriction (SAAR) compared with control (Con) diet for seven days. (B) Normalized count values of Vegfb in EDL and soleus from bulk RNA sequencing ( n = 6) of male mice given ad libitum access to SAAR versus Con diet on day seven using transcriptomic dataset presented in . (C) Normalized count values of Flt1 in EDL and soleus from bulk RNA sequencing ( n = 6) of male mice given ad libitum access to SAAR versus Con diet on day seven using transcriptomic dataset presented in . (D) Experimental set up and color scheme used in E, 5F, C, and S5D. (E) Percent change in body weight ( n = 16–24) of male mice treated with vehicle (veh) or axitinib via oral gavage in combination with ad libitum access to SAAR versus Con diet after seven days. (F) Distance ran during a one-time maximal endurance test ( n = 16–24) of male mice treated with veh or axitinib via oral gavage in combination with ad libitum access to SAAR versus Con diet for seven days. (G) Representative fluorescence images of IB4 (white) staining in EDL cross sections of mice fed a Con or SAAR Diet, co-treated with veh or axitinib via oral gavage (scale bar, 400 μm) for seven days (n = 5–8). (H) Normalized count values of Flt1 in EDL treated with either veh or axitinib from bulk RNA sequencing ( n = 5) of male mice given ad libitum access to SAAR versus Con diet on day seven. (I) Normalized count values of Kdr in EDL treated with either veh or axitinib from bulk RNA sequencing ( n = 5) of male mice given ad libitum access to SAAR versus Con diet on day seven. (J) Experimental set up and color scheme used in K, 5L, J, and S5K. (K) Percent change in body weight (n = 8–10) of male mice treated with IgG or DC101 via i.p. injection every other day in combination with ad libitum access to SAAR versus Con diet after seven days. (L) Distance ran during a one-time maximal endurance test ( n = 12–16) of male mice treated with IgG or DC101 via i.p. injection every other day in combination with ad libitum access to SAAR versus Con diet on day seven. A-C, E, G-I, represent data from mice that were not subjected to endurance running; F, K, and L represent data from mice subjected to maximal endurance testing. All data are shown as mean, and error bars indicate SD unless otherwise noted; p values indicate the significance of the difference by two-way ANOVA with Sidak multiple comparisons test between diets and treatment; significance is determined by p < 0.05. See also and .

Article Snippet: Mouse (IgG2b) Myosin Heavy Chain Type I antibody (BA-F8) , DSHB , Cat# BA-F8; RRID: AB_10572253.

Techniques: Inhibition, Muscles, Control, RNA Sequencing, Fluorescence, Staining, Injection