Journal: Nature Communications

Article Title: FANCD2 restrains fork progression and prevents fragility at early origins upon re-replication

doi: 10.1038/s41467-026-68966-4

Figure Lengend Snippet: a Schematic of the screen; hTERT-RPE1 cells were transfected with control or Geminin siRNAs and the designed siRNA library prior to EdU pulse, fixation and immunostaining for γH2AX. Nuclei were also stained with DAPI. Analyses were based on mean intensities and robust z-score transformation in respect to negative controls. b Left: Mean robust z-scores of γΗ2ΑΧ intensities and total cell counts for Geminin-depleted cells. Right: Top ranked genes in the Geminin-depleted cell model and corresponding values in control cells for γΗ2ΑΧ nuclear intensities and total cell counts. c Top: Immunoblot detection of indicated proteins in hTERT-RPE1 cells transfected with indicated siRNAs. α-Tubulin, loading control. Bottom: Images and quantification of transfected hTERT-RPE1 cells stained for γH2AX (left) and 53BP1 (right). For γH2AX intensities, data pooled from n = 3 biological replicates (> 250 cells per condition and replicate); two-tailed Mann-Whitney U test, **** p < 0.0001, ns = not significant. For cells with >10 53BP1 foci, plot shows mean ± SD of n = 4 biological replicates (> 250 cells per condition and replicate); two-tailed unpaired t-test, *** p = 0.0007, ** p = 0.0054, * p = 0.0120, ns = not significant. d Schematic of the workflow, representative images and quantification of long-term clonogenic assays in hTERT-RPE1 cells transfected with indicated siRNAs. Plot shows mean ± SD of n = 3 biological replicates; two-tailed unpaired t-test, **** p < 0.0001, *** p = 0.0003, * p = bottom to top; 0.0177, 0.0158. DNA staining, Hoechst. Scale bar, 10 μm. siCtrl, negative siRNA control; siGMNN, siRNA targeting Geminin; siFANCD2, siRNA targeting FANCD2. Source data are provided as a Source Data file.

Article Snippet: To generate a U2OS cell line with doxycycline-inducible depletion of Geminin, shRNA sequences targeting human Geminin (GMNN) were cloned into the Tet-pLKO-puro “all-in-one” lentiviral system for tetracycline-inducible shRNA expression (Addgene #21915). shRNA sequences were selected from the RNAi Consortium collection (MISSION®, Sigma-Aldrich) and synthesized oligonucleotides for each shRNA were annealed and ligated into the Tet-pLKO-puro vector following enzimatic digestion followed by transformation into competent bacteria.

Techniques: Transfection, Control, Immunostaining, Staining, Transformation Assay, Western Blot, Two Tailed Test, MANN-WHITNEY

Journal: Poultry Science

Article Title: Prevalence of pathogenic bacteria and their antimicrobial patterns analysis of clinical samples from free-range chickens raised in forest farms in Zhouqu county of Gansu Province, China

doi: 10.1016/j.psj.2025.105859

Figure Lengend Snippet: List of antimicrobials included in susceptibility analysis.

Article Snippet: 6. Tetracycline (TET) , 30 , Liofilchem. Srl, Italy.

Techniques: