Journal: iScience

Article Title: Activation of SNr GABA neurons drives liver-brain-eye axis dysfunction in hepatic encephalopathy

doi: 10.1016/j.isci.2026.114720

Figure Lengend Snippet: Screening strategy for identifying brain nuclei associated with HE-related visual impairment using SNr as a seed region (A) Schematic workflow of SNr viral injection in C57BL/6J and GAD2-Cre mice. Virus injection was performed on day 1, TAA was injected daily on days 19–21, and samples were collected on day 22. (B) Diagram of left SNr viral injection in C57BL/6J mice. (C) Diagram showing projections from the SNr to downstream regions in C57BL/6J mice (∗ indicates downstream projection areas). (D) “+” indicates the presence of SNr-to-region projections; “−” indicates the absence of such projections. (E) Schematic of specific downstream nuclei receiving projections from the SNr in C57BL/6J mice. (F) Among downstream nuclei, 10 were positive only in the control group, 6 were positive only in the AHE group, and 43 were positive in both groups. (G) Workflow of SNr viral injection in c-Fos-Cre ERT2 mice. Virus injection was performed on day 0, tamoxifen was injected daily on days 8–14, TAA was injected daily on days 19–21, and samples were collected on day 22. (H) Diagram of left SNr viral injection in c-Fos-Cre ERT2 mice. (I) Diagram showing projections from the SNr to downstream activated regions in c-Fos-Cre ERT2 mice (∗ indicates downstream activated projection areas). (J) Schematic of specific downstream regions receiving projections from the SNr in c-Fos-Cre ERT2 mice. (K) Among downstream nuclei, 2 were positive only in the control group, 17 were positive only in the AHE group, and 19 were positive in both groups. (L) Diagram of left SNr viral injection in GAD2-Cre mice. (M) Diagram showing downstream nuclei receiving projections from mSNr in GAD2-Cre mice (∗ indicates downstream projection nuclei). (N) Schematic of specific downstream regions receiving projections from mSNr in GAD2-Cre mice. (O) Among activated downstream nuclei, none were positive only in the control group, 9 were positive only in the AHE group, and 5 were positive in both groups. (P) The 17 nuclei activated only in the AHE condition were selected for further analysis. (Q) Intersection of the 9 nuclei activated only in the AHE condition in GAD2-Cre mice and the 17 nuclei activated only in the AHE condition in TRAP mice resulted in the selection of 5 nuclei. (R) These 5 nuclei represent the mSNr GAD2+ regions activated under HE conditions. (S) Schematic diagram of SNr viral injection and AHE model construction in C57BL/6J mice. Viral injection was performed on day 1, followed by TAA injections on days 19–21 for 3 consecutive days, and tissue collection on day 22. (T) Diagram of SNr viral injection site (scale bar, 1 mm). (U) Projection maps of SNr downstream nuclei at bregma = −4.35 mm and interaural = −0.56 mm in control and AHE groups (scale bars, 1 mm). (V) Projection maps of SNr downstream nuclei at bregma = −1.58 mm and interaural = −2.22 mm in control and AHE groups (scale bars, 1 mm). (W) Projection maps of SNr downstream nuclei at bregma = −3.28 mm and interaural = 0.52 mm in control and AHE groups (scale bars, 1 mm). (X) Projection maps of SNr downstream nuclei at bregma = −3.8 mm and interaural = 0.00 mm in control and AHE groups (scale bars, 1 mm). (Y) Projection maps of SNr downstream nuclei at bregma = −4.16 mm and interaural = −0.36 mm in control and AHE groups (scale bars, 1 mm). (Z) Projection maps of SNr downstream nuclei at bregma = −4.90 mm and interaural = −1.60 mm in control and AHE groups (scale bars, 1 mm).

Article Snippet: The c-Fos-Cre ERT2 transgenic mice (designated as TRAP) were obtained from Jackson Laboratory (FOS tm2.1 (icre/ERT2) Luo/J ; stock number #030323), aged 6–8 weeks.

Techniques: Injection, Virus, Control, Selection

Journal: bioRxiv

Article Title: OSTM1 is a ubiquitin E3 ligase that suppresses B-cell malignancy by activating the cAMP/PKA/CREB pathway

doi: 10.64898/2026.01.23.701155

Figure Lengend Snippet: Ostm1 lox/lox mice were bred to Cd19 -Cre and Cdkn2a lox/lox mice. A ) Kaplan-Meier surivival survival curves of littermate control wild-type (LMC-WT), Ostm1 -/- (O -/- ), Cdkn2a -/- (C -/- ), Ostm1 +/- ; Cdkn2a -/- (O +/- ;C -/- ), and Ostm1 -/- ; Cdkn2a -/- (DKO) mice. ( B ) Spleen and liver weight at the time of end-point O +/- ;C -/- or DKO mice, comparing with the age-and gender-matched WT and single knockout (SKO) mice. ( C ) Spleen images and histopathological features of indicated genotypes analyzed by H&E and IHC staining. ( D ) Incidence of BCL in the spleen, lymph nodes (LNs), and bone marrow (BM), based on clonal abnormal B-cell populations. ( E ) The percentage of normal follicular B cells (FO; B220 + CD3 - CD23 + CD21 int ), marginal zone B cells (MZ; B220 + CD3 - CD23 int CD21 + ), and double negative B cells (B220 + CD3 - CD21 - CD23 - ; activated/immature/malignant B cells) in the spleen of the mice analyzed in (A). **p<0.01; ***p<0.001 as determined by ANOVA. (F) Representative transplantation of DKO splenic BCL into nude recipient mice. Splenocytes (1×10 6 ) isolated from DKO mice (ID: 4491 or 4531) were injected intraperitoneally (i.p.) into each nude recipient mouse. At 7-9 weeks post-transplantation, the spleen, LN, and BM cells were harvested and analyzed by flow cytometry (FACS). FACS profiles of primary DKO splenocytes are shown for comparison. (G-J) Mice of the indicated genotypes were harvested at the endpoint of the DKO cohort (around 31-week-old). Bulk RNA-seq was performed on purified splenic B-cells. (G) Heatmap showing genes that were markedly down-regulated (Cluster A) and upregulated (Cluster B) in DKO mice. (H and I) KEGG pathway enrichment analysis of genes in Cluster A (H) and Cluster B (I) was performed. The osteoclast differentiation pathway is highlighted. (J) Disease-associated gene enrichment analysis of Cluster A (down-regulated genes in DKO mice). B-cell lymphomagenesis-related pathways are highlighted in red.

Article Snippet: Cdkn2a flox/flox mice (B6.129S1- Cdkn2a /J, Strain #: 023323), and Cd19 +/Cre mice (B6.129P2(C)- Cd19 tm1(cre)Cgn /J) were purchased from the Jackson Laboratory.

Techniques: Control, Knock-Out, Immunohistochemistry, Transplantation Assay, Isolation, Injection, Flow Cytometry, Comparison, RNA Sequencing, Purification

Journal: bioRxiv

Article Title: Cell type-specific proximity labeling of organ secretomes reveals energy balance-dependent proteomic remodeling

doi: 10.64898/2026.01.11.698831

Figure Lengend Snippet: (A) Schematic of LoxP-flanked TurboID-KDEL construct and ER-localized TurboID proximity labeling system. (B) Experimental design for comparison of three tissues under basal conditions. (C) Western blot showing TurboID-catalyzed biotinylation of inguinal adipose tissue and plasma from biotin-treated Adipo-TurboID KDEL mice, as well as V5 tag indicative of TurboID expression. (D) Immunohistochemistry images of inguinal white adipose tissue (iWAT), liver, and spleen tissue in TurboID KDEL mice with and without expression of Adiponectin-Cre, Albumin-Cre or CD19-Cre. Biotinylation is shown in red (streptavidin for iWAT, neutravidin for liver and spleen). Scale bar 50 µm. (E) Workflow for tissue processing, affinity purification, and analysis by Tandem-Mass-Tag (TMT) mass spectrometry. (F) Pathway analysis illustrating predicted subcellular localization of proteins enriched in basal Adipo-TurboID KDEL Cre+ relative to Cre- samples. Dashed line indicates false discovery rate (FDR) of 5%. Gene Ontology term enrichment analysis was performed with GO Slim Cellular Component with the full mouse genome used as the background list. (G) Protein abundance plots highlighting cell-type-specific markers expressed in liver, iWAT, and spleen, respectively.

Article Snippet: CD19-Cre (B6.129P2(C)-Cd19tm 1(cre)Cgn /J, Jackson Laboratory 006785), Adiponectin-Cre (B6.FVB-Tg(Adipoq-cre)1Evdr/J, Jackson Laboratory 028020), and LSL-TurboID-KDEL, which was generated in the transgenic core facility at Rockefeller University.

Techniques: Construct, Labeling, Comparison, Western Blot, Clinical Proteomics, Expressing, Immunohistochemistry, Affinity Purification, Mass Spectrometry, Quantitative Proteomics