Journal: Nature Communications

Article Title: Tuning intracellular immunity by Nodamura virus B2 protein enhances self-amplifying RNA activity

doi: 10.1038/s41467-026-77816-2

Figure Lengend Snippet: A Representative contour plots of GFP + HeLa cells 24 h post-transfection with CHIKV-GFP-STOP (blue) or -B2 WT (red) saRNA. Gate indicates the GFP high population used for MFI analysis in ( B , C ). B GFP high MFI monitored by flow cytometry in HeLa cells transfected with CHIKV-GFP-STOP (blue) or -B2 WT (red) at 16, 24 and 48 h post-transfection. C Area under the curve for GFP high MFI values in ( B ). Mean ± s.d., n = 3 independent experiments with each point the mean of technical replicates. D , E Representative images of GFP signal in HeLa cells 24 h post-transfection of CHIKV-GFP-B2 WT or -B2 R59Q in the presence of DMSO ( D ) or ruxolitinib ( E ). GFP (green), DAPI (blue), scale bar: 100 μm. F , G GFP high MFI ( F ) or percentage of GFP + cells ( G ) in HeLa cells transfected with CHIKV-GFP-STOP (blue), -B2 WT (red) or -B2 R59Q (yellow) saRNA in the presence of DMSO, ruxolitinib (RUXO) or recombinant universal IFN-I (IFNα-A/D) 24 h post-transfection. H, I HeLa cells transfected with CHIKV-GFP-STOP (blue), -B2 WT (red), -B2 R59Q (yellow) saRNA or poly(I:C) (100 ng for 24 h). Mean ± s.d., n = 3 independent experiments with each point representing the mean of technical replicates. H RT-qPCR analysis of mRNA specific to GFP (left) or nsP1 (right) mRNA 24 h post-transfection. I RT-qPCR of ISGs IFIT1 (left), IFIT2 (middle) or RIG-I (right) mRNAs. J GFP high MFI of WT HEK293 cells or RIG-I -/- MDA5 -/- cells 24 h post-transfection with of CHIKV-GFP-B2 WT (red) or -B2 R59Q (yellow). K WT and RIG-I -/- MDA5 -/- HEK293 cells were treated as in ( J ) and IFIT1 (left) and IFIT2 (right) mRNAs were analysed by RT-qPCR. Data are mean ± s.d. from a representative of two biological replicate experiments. Connected dots correspond to values from the same biological replicate. Statistical analyses were performed using a two-tailed ratio paired t-test ( C ) and a two-way ANOVA with Šídák’s multiple comparisons test ( B ), Dunnett’s multiple comparisons test ( F , G ) or Fisher’s LSD test ( J ). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns not significant. arb. units: arbitrary units. Exact p values for ( B , C , F , G , J ) and source data are provided the Source Data file.

Article Snippet: After transfection, cells were transferred to the appropriate well format with complete medium or, where indicated, complete medium containing PKR inhibitor C16 (527450, Sigma-Aldrich), or 500 nM of JAK inhibitor Ruxolitinib (tlrl-rux, Invivogen), or 500 UI mL −1 of universal type I IFN (human IFN alpah A/D) (11200, PBL assay Science).

Techniques: Transfection, Analysis, Flow Cytometry, Recombinant, Quantitative RT-PCR, Two Tailed Test

Journal: Stem Cell Reports

Article Title: CCL5 exposure promotes acute and transient megakaryopoiesis and hematopoietic progenitor expansion

doi: 10.1016/j.stemcr.2026.103014

Figure Lengend Snippet: CCL5 administration induces transcriptional changes in megakaryocytes (A) Mice were treated with CCL5 as previously described, and bone marrow MKs were isolated after 24 h following lineage depletion and enrichment by flow cytometry sorting. RNA was extracted using TRIzol purification, and RNA sequencing was performed (4n vs. 4n). (B) Volcano plot shows genes that are significantly up- and down-regulated. The points highlighted in purple are genes that have padj <0.05 and a log2-fold change >1. Points in blue have a padj <0.05 and a log2-fold change <1. Gray points are non-significant. The dashed lines correspond to the cutoff values of log2 fold-change and padj chosen. (C) Differentially expressed genes associated with mitochondrial metabolism. Expression values are presented in log 2 scale, and statistical significance was determined using padj. N represents a pool of 2 mice. (D) Male mice received retro-orbital injections of CCL5 (0.05 mg/kg) or vehicle ( Ctrl ). After 24 h, MKs were isolated by size exclusion, lysed, and analyzed using a RayBio Mouse JAK/STAT Pathway Phosphorylation Array. Signal intensity was quantified with Fiji. N represents a pool of 5 mice per group. (E) Representative histograms of TMRM in MKs by flow cytometry after CCL5 administration (Gray: FMO; Black: Ctrl; Red: CCL5). (F) MKs were identified by flow cytometry, and the mean fluorescence intensity of TMRM was quantified ( n = 4–5). Data are presented as mean ± SD. Statistical analyses were performed using an unpaired, two-tailed Student’s t test. N represents an individual mouse. Illustrations were created with BioRender. padj, adjusted p value.

Article Snippet: After 24 h, MKs were isolated by size exclusion, lysed, and analyzed using a RayBio Mouse JAK/STAT Pathway Phosphorylation Array.

Techniques: Isolation, Flow Cytometry, Purification, RNA Sequencing, Expressing, Phospho-proteomics, Fluorescence, Two Tailed Test

Journal: Acta Pharmaceutica Sinica. B

Article Title: Targeting NAT10 with Remodelin reshapes the pro-fibrotic microenvironment in renal fibrogenesis via inhibiting exosome oversecretion from tubular cells

doi: 10.1016/j.apsb.2026.05.020

Figure Lengend Snippet: Targeting acetyltransferase NAT10 with Remodelin breaks the TEC-fibroblast pro-fibrotic chain (A) The graph indicating the selection strategy of screen based on in vitro model of TEC-fibroblast communication with epigenetic inhibitor compounds. (B) The heatmap of viability of NRK-49F cells incubated with CM from HK-2 cells with indicated treatments respectively (left panel). The dots indicated the average fold of viability of NRK-49F cells incubated with CM from HK-2 cells treated with TGF- β and epigenetic inhibitors, in relative to the NRK-49F cells incubated with CM from HK-2 cells treated with TGF- β and DMSO (right panel). The red dots indicated the selected inhibitors for further screen. (C) The heatmap of viability of NRK-49F cells incubated with CM from HK-2 cells with indicated treatments, respectively. (D) The heatmap (left) and table (right) demonstrating the details of 13 effective epigenetic inhibitors. (E) The immunoblot analysis of NAT10 in kidneys from sham mice ( n = 5) and UUO mice ( n = 5). (F) The dot blot analysis of ac4C in kidneys from sham mice ( n = 2) and UUO mice ( n = 2). (G) The immunohistochemistry staining of NAT10 in kidneys from sham mice and UUO mice. (H) The immunofluorescence staining of NAT10 in kidneys from sham mice and UUO mice. LTL was used as marker of proximal tubular cells. (I) The immunoblot analysis of NAT10 in kidneys from CKD patients ( n = 18) and non-CKD control ( n = 5). (J) The dot blot analysis of ac4C in kidneys from CKD patients ( n = 18) and non-CKD control ( n = 5). Data are presented as mean ± SD. Student's t test was used for comparison between two groups and one-way ANOVA was used for comparison between multiple groups; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

Article Snippet: HK-2 cells were seeded in 96-well plates at the density of 10,000 cells per well, and then treated with TGF- β (5 ng/mL) in the presence of each inhibitor (1 or 5 μmol/L) of Epigenetic Compound Library (TargetMol, MA, USA).

Techniques: Selection, In Vitro, Incubation, Western Blot, Analysis, Dot Blot, Immunohistochemistry, Staining, Immunofluorescence, Marker, Control, Comparison