Journal: bioRxiv

Article Title: High avidity phase-separated RNA-protein sialogranules sense lectins and inhibit influenza infection

doi: 10.64898/2026.02.22.707264

Figure Lengend Snippet: (A) Predicted AlphaFold structure for LAMP1-mCherry-tdPCP. Black, red, and green are LAMP1, mCherry, and tdPCP domains, respectively. Putative sialylation sites are marked by light-blue shading (asparagine residues in an N-X-S/T motif), yellow (serine residue), and magenta (threonine residue). (B) Typical field-of-view of LAMP1 granule (top) and the MABs (bottom) divided into individual channels. (Top-left) protein (mCherry). (Top-right) SNA (AF488). (Bottom-left) slncRNA (AF405). (Bottom-right) Merged panel with all three images overlayed. (Top-set) No SNA added (scalebar: 5µm FoV, 1µm enlargement). (Bottom-set) High SNA concentration, corresponding to the 2mg/ml stock (scalebar: 5µm). (C) Box-plot distributions for D RNA for each candidate sialoprotein. The mean of each distribution is marked by dark red X. Each red dot represents one single mCherry-positive event for which the D RNA was calculated. Low and high SNA concentrations were set at below 1µg/ml (molar ratio of ∼1:50 SNA:sialoproteins) and above 50µg/ml (molar ratio of ∼3:2 SNA:sialoproteins), respectively. The student’s t-test was applied to ascertain statistical significance. (D) Δ D as a function of putative sialylated sited. Δ D is defined as the difference between the mean D RNA values for high and low SNA concentrations obtained for each protein in panel C. Color and size of circle correspond to the student’s t-test p-value computed for each panel in . We note that NXST1m and NXST2m appear to have nearly the same Δ D value and have the same number of putative sialylated sites.

Article Snippet: For fluorescently labeled RNA, 1μg of the restriction product was used as template for in vitro transcription using HighYield T7 Alexafluor-405 (AF405) RNA labeling kit (Jena Bioscience, cat. RNT-101-AF405), according to the manufacturer’s instructions.

Techniques: Residue, Concentration Assay

Journal: bioRxiv

Article Title: High avidity phase-separated RNA-protein sialogranules sense lectins and inhibit influenza infection

doi: 10.64898/2026.02.22.707264

Figure Lengend Snippet: (A) Typical field-of-view of SNA-LAMP1 MABs divided into individual channels. (Top-left) protein (mCherry). (Top-right) SNA (AF488). (Bottom-left) slncRNA (AF405). (Bottom-right) Merged panel with all three images overlayed. Scalebar: 5µm. (B) Close-up on the two features highlighted in panel A displayed via the same three separated channels and merged image. Scalebar: 5μm. (C) Left: diagram of event classification: a cluster of mCherry-positive events is colocalized with clusters of FITC- or AF405-positive pixels if the overlap is at least 75% and 15%, respectively. Triple labelled events are counted towards the SNA-binding bin. Right: LAMP1 dose-response curves (right) for sialogranule (blue) and MAB (green) event frequency produced based on the decision diagram. Percentages represent the goodness of fit to a Hill curve (B – blue, G – green). The dose-response curves for the other proteins is shown in Figure S3. (D) Dose responses (see Figure S3B for all proteins) displayed as a normalized heatmap for SNA binding (top) and RNA displacement (bottom). Each row was normalized by its maximal value. (E) The change in Gibbs’ free energy (ΔΔ G ) computed based on the event frequency K d (see methods) for both the granules and MABs is dependent on the number of putative sialylated sites. A logarithmic fit is demonstrated between the number of putative sialylated sites and either the accumulation MAB (left, R 2 =0.88) or the reduction of sialogranule events (right, R 2 =0.94).

Article Snippet: For fluorescently labeled RNA, 1μg of the restriction product was used as template for in vitro transcription using HighYield T7 Alexafluor-405 (AF405) RNA labeling kit (Jena Bioscience, cat. RNT-101-AF405), according to the manufacturer’s instructions.

Techniques: Binding Assay, Produced

Journal: bioRxiv

Article Title: Restoring Shugoshin 1 reduces chromosome errors in human eggs

doi: 10.64898/2026.01.08.698387

Figure Lengend Snippet: (A) Representative immunofluorescence images showing the localization nascent transcripts (EU, green) inside the nucleus (blue) of early GV oocytes. Images show EU levels in negative control (no EU), control (with EU), upon treatment with α-amanitin, and triptolide. Scale bar: 10 µm. (B) Frequency of PSSC with α-amanitin measured by scoring the percentage of eggs with PSSC, similar to levels observed with triptolide . Plots show number of eggs analyzed on top. Statistical significance is measured by Fisher’s exact test, ns = not significant. (C) Representative time-lapse images showing the localization of MajSat RNA in GV and MI oocytes and MII eggs upon microinjection of GV oocytes with UTP-X-Cy3 labeled MajSat (top panel) and control (bottom panel) RNA. MajSat RNA localizes to the pericentromeres during metaphase I and metaphase II stages along with foci present in the cytoplasm. Cy3-labeled control RNA localizes only in the cytoplasm and not on chromosomes. MajSat and control RNA (green), chromosomes (H2B-SNAP, blue) are shown. Scale bars: 10 µm. (D) Co-localization of MajSat RNA and SGO1 observed on metaphase I chromosome spreads upon performing RNA FISH with immunofluorescence. Scale bar: 10 µm.

Article Snippet: The HighYield T7 Cy3 RNA Labelling Kit (Jena Bioscience, #RNT-101-CY3) was used to label amplified sequences with Cy3-labelled UTP (UTP-X-Cy3, 35%), followed by DNA removal using TurboTM DNAse (ThermoFisher).

Techniques: Immunofluorescence, Negative Control, Control, Microinjection, Labeling

Journal: Fundamental Research

Article Title: Disruption of the nucleocapsid-RNA condensation by punicalagin is a broad-spectrum antiviral approach

doi: 10.1016/j.fmre.2025.07.007

Figure Lengend Snippet: Punicalagin binds to N and inhibits SARS-CoV-2 replication in cells. (a and b) The inhibitory effect of punicalagin with indicated concentrations on the N condensates under SARS-CoV-2 infection. Representative immunofluorescent images were shown (a). The foci number per cell was quantified (b). 3D images were obtained by a Zeiss LSM 880 confocal microscope and reconstituted by Volocity 6.1.1 . Scale, 1 unit = 3.62 µm. (c) The inhibitory effect of punicalagin with indicated concentrations on the replication of SARS-CoV-2. IC 50 was calculated. (d) The effect of punicalagin on viral entry. A549-hACE2 cells were infected with pSARS2-VSV-Nluc pseudoviruses, following a 1-h pretreatment of punicalagin (10 µM). Luminescence was detected to quantify the entry of virus. (e) The effect of punicalagin on RdRp activity. A549-hACE2 cells were transduced with pSARS2-Replicon-Nluc, following a 1-h pretreatment of punicalagin (10 µM). Luminescence was detected to quantify the genome replication of virus. (f) The effect of punicalagin on 3CL pro activity. 3CL pro recombinant proteins were incubated with punicalagin, and the activity was then quantified through the optical density of cleaved substrate. (g) EMSA showed the RNA binding ability of N protein, Kaempferol and punicalagin. Cy5-labeled vRNA (60 nt) was used as RNA probe. (h) The effect of punicalagin on the RNA-binding capacity of N-mEGFP protein was determined by EMSA. Cy5-labeled vRNA (60 nt) was used as RNA probe. (i) Immunoblot analysis of punicalagin-N interaction. Pull-down assay was performed by punicalagin-conjugated agarose beads incubated with the lysate of A549 cells transfected with pLVX-N-Flag. Indicated antibodies were used. (j) Schematic drawing of the N protein. (k) Punicalagin-conjugated agarose beads incubated with lysate of HEK293T cells transfected with indicated pLVX-N-Flag truncations, and the punicalagin-N truncations interaction was analyzed by immunoprecipitation. Pun, 0 µM, PBS (a-c, h). Negative control (Ctrl), PBS (a-c), DMSO (d, e and f) and XB buffer (g and h). Statistical analysis was performed using a two-tailed unpaired Student’s t -test (b). IC 50 values were shown as mean ± s .d., calculated by nonlinear regression using a four-parameter logistic model (c). Data are presented as mean ± s .e.m. (b), or mean ± s .d. (c-e and f). Sample sizes were n = 60 cells (b), 3 biologically independent experiments (c and f), and 6 biologically independent replicates (d and e). The conclusions presented were repeated at least three times. Source data are provided as a Source Data file.

Article Snippet: Full-length SARS-CoV-2 3′ UTR was in vitro transcribed and labeled with HyperScribe T7 High Yield Cyanine 5 (Cy5) RNA Labeling Kit (E2040S, NEB).

Techniques: Infection, Microscopy, Virus, Activity Assay, Transduction, Recombinant, Incubation, RNA Binding Assay, Labeling, Western Blot, Pull Down Assay, Transfection, Immunoprecipitation, Negative Control, Two Tailed Test