Journal: Development (Cambridge, England)

Article Title: CSF1R + macrophage and osteoclast depletion impairs neural crest proliferation and craniofacial morphogenesis

doi: 10.1242/dev.205423

Figure Lengend Snippet: Gestational exposure to PLX5622 significantly depletes CSF1R-expressing cells. (A) Schematic illustrating collection of Csf1r EGFP + cells for flow cytometry. (B) Quantification of EGFP + cells from Csf1r EGFP craniofacial tissues ( n =3-10 embryos per sex/treatment/time-point from two to four dams). (C-W) Immunofluorescence images and quantification of CSF1R + and Csf1r EGFP+ cells in and around the E15.5 nasal septum (C-E), Meckel's cartilage (F-H), ear (I-K), maxillary incisor (L-N), eye (O-Q), trigeminal (R-T) and tongue (U-W). Arrows mark CSF1R and Csf1r EGFP double-positive cells ( n =3 embryos per sex/treatment from two or three dams). c.d., cochlear duct; d.p., dental papilla; e.k., enamel knot; ey, eye; l.s.c., lateral semicircular canal; m.c., Meckel's cartilage; n.s., nasal septum; s.r., stellate reticulum; tg, tongue; ut, utricle. Blue dots represent male and pink dots represent female. Counts represent mean±s.e.m. and were analyzed by a two-way ANOVA with Tukey's post-hoc test.

Article Snippet: Csf1r EGFP mice (JAX: 005304; RRID: IMSR_JAX:005304; The Jackson Laboratory) were crossed to C57BL/6 mice to generate heterozygous embryos for flow cytometry and immunofluorescence experiments.

Techniques: Expressing, Flow Cytometry, Immunofluorescence

Journal: Development (Cambridge, England)

Article Title: CSF1R + macrophage and osteoclast depletion impairs neural crest proliferation and craniofacial morphogenesis

doi: 10.1242/dev.205423

Figure Lengend Snippet: Prenatal exposure to PLX5622 disrupts osteoclast development and function. (A-D′) Csf1r EGFP + osteoclasts (arrows) in the E15.5 premaxilla (A-B′) and maxilla (C-D′). (E-I) Immunofluorescence images (E-H) and quantification (I,I′) of multinucleated CTSK/ Csf1r EGFP double-positive osteoclasts (arrows) (I) and Csf1r EGFP+ single-positive osteoclasts (I′) in the E15.5 mandible. (J-S′) TRAP staining (arrows) in E15.5 premaxilla (J-K′), mandible (L-M′), maxilla (N-O′), frontal (P-Q′) and basioccipital (R-S′) bones. (T-V) Quantification of total TRAP + area in premaxilla (T), mandible (U) and maxilla (V). n =3 embryos per sex/treatment from two or three dams. bo, basioccipital; fr, frontal; m.c, Meckel's cartilage; md, mandible; mx, maxilla pmx, premaxilla. Counts represent mean±s.e.m. and were analyzed by an ART ANOVA (I,I′) or a two-way ANOVA (T-V) with Tukey's post-hoc test. A′-F′ show magnifications of the respective boxed areas in A-F.

Article Snippet: Csf1r EGFP mice (JAX: 005304; RRID: IMSR_JAX:005304; The Jackson Laboratory) were crossed to C57BL/6 mice to generate heterozygous embryos for flow cytometry and immunofluorescence experiments.

Techniques: Immunofluorescence, Staining

Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: Schematic illustration of in vivo tumor immunotherapy enhanced by mRNA/HNPs through intravenous injection. H18 lipid, DOPE, cholesterol, DMG-PEG 2000 and mRNA were mixed to form mRNA/H 18 NPs with the special multilamellar concentric nanostructures. Following intravenous administration, mRNA/H 18 NPs demonstrated preferential adsorption of complement C3 proteins to form a characteristic protein corona, resulting in specific mRNA transfection in the spleen, especially in splenic dendritic cells. When encapsulating tumor antigen-encoding mRNA, the mRNA/H 18 NPs achieved precise transfection of the antigen mRNA in splenic dendritic cells. This targeted delivery stimulated dendritic cell maturation and subsequent antigen presentation, initiating robust T cell priming. The activated antigen-specific cytotoxic T lymphocytes then infiltrated into tumor tissues, ultimately inducing tumor cell elimination.

Article Snippet: Luciferase mRNA, OVA mRNA, and EGFP mRNA were purchased from Novoprotein (Suzhou, China).

Techniques: In Vivo, Injection, Adsorption, Transfection, Immunopeptidomics

Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: Preparation and Characterization of Optimal mRNA/H 18 NPs. (A) Schematic illustration of mRNA/H 18 NPs preparation. (B) The size distribution and (C) zeta potential of optimized mRNA/H 18 NPs. (D) The apparent p K a of mRNA/H 18 NPs. (E) Cryo-EM image of optimized mRNA/H 18 NPs. Scale bar = 50 nm. (F) Representative image and percentage of bioluminescence in major organs of mice following intravenous injection of mLuc/H 18 NPs. (G)-(I) Stability test for mRNA/H 18 NPs. (G) Size and PDI of mRNA/H 18 NPs when stored at 4 °C for different days (0, 3, 5, 7). (H) Left: Total bioluminescence flux in the spleen of mice 6 h after intravenous injection of mLuc/H 18 NPs stored at 4 °C for different days (0, 3, 5, 7). Right: Representative bioluminescence images of major organs of mice 6 h after intravenous injection of different mLuc/H 18 NPs stored at 4 °C for different days (0, 3, 5, 7). (I) Size and PDI of mRNA/H 18 NPs when diluted with PBS by different times. Data were shown as mean ± SD (n = 3).

Article Snippet: Luciferase mRNA, OVA mRNA, and EGFP mRNA were purchased from Novoprotein (Suzhou, China).

Techniques: Zeta Potential Analyzer, Cryo-EM Sample Prep, Injection

Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: In vivo splenic DC-specific transfection of mRNA/H 18 NPs and in vitro protein corona analysis of mRNA/H 18 NPs. (A) EGFP protein expression in splenic cell subsets of C57BL/6J mice 24 h post intravenous injection of different formulations. (B) The top 5 most abundant plasma proteins adsorbed on mRNA/H 18 NPs (C3: Complement C3; Ighm: Immunoglobulin heavy constant mu; Hbat1: Alpha-globin; Itih4: Inter alpha-trypsin inhibitor, heavy chain 4; Cnn2: Calponin). (C) Heatmap plot of major proteins in the protein corona adsorbed on mRNA/MC3-LNPs and mRNA/H 18 NPs. PBS group was used as a negative control. (D) Quantification of major adsorbed protein categories of different formulations. (E) Complement C3 abundance in protein corona adsorbed on mRNA/MC3-LNPs and mRNA/H 18 NPs. (F) Bioluminescence images of major organs and (G) Quantification of total bioluminescence flux in the spleen from C57BL/6J mice 6 h after intravenous injection of mLuc/H 18 NPs (mLuc dose of 0.25 mg kg −1 ). Mice were pre-treated with cobra venom factor (CVF) or PBS. (H) Fluorescence quantification of Cy5 mRNA delivered by uncoated or complement C3-coated Cy5-mRNA/H 18 NPs in BMDCs. BMDCs were pre-incubated with anti-CD11b (CR3) or anti-IgG blocking antibody. (I) Bioluminescence intensity of luciferase protein translated from mRNA delivered by uncoated or complement C3-coated mLuc/H 18 NPs in BMDCs. BMDCs were pre-incubated with anti-CD11b (CR3) or anti-IgG blocking antibody. Data were shown as mean ± SD (n = 3).

Article Snippet: Luciferase mRNA, OVA mRNA, and EGFP mRNA were purchased from Novoprotein (Suzhou, China).

Techniques: In Vivo, Transfection, In Vitro, Expressing, Injection, Clinical Proteomics, Negative Control, Combined Bisulfite Restriction Analysis Assay, Fluorescence, Incubation, Blocking Assay, Luciferase

Journal: STAR Protocols

Article Title: Protocol for an in vivo CRISPR screen for germinal center B cells in mice using ecotropic retrovirus

doi: 10.1016/j.xpro.2026.104586

Figure Lengend Snippet: Immunization experiment scheme for in vivo screen Host mice (IOMA gl) were immunized with OVA in alum. At week 2 after immunization, B cells from B1-8 hi , LSL-Cas9, AID-Cre, Kappa KO animals were transduced with pooled sgRNA library retrovirus and adoptively transferred to the hosts. Subsequently, the host animals were boosted with NP-OVA in alum at day 0 and 2 and administered with anti-DEC205-OVA mAb at day 6.5. Spleens were harvested and processed for cell sorting at day 10.

Article Snippet: Mouse: 6-weeks-old, sex-matched LSL-Cas9: B6J.129(B6N)-Gt(ROSA)26Sortm1(CAG-cas9∗,-EGFP)Fezh/J , The Jackson Laboratory , cat# 026175.

Techniques: In Vivo, Transduction, FACS

Journal: International Journal of Pharmaceutics: X

Article Title: A pH-responsive dual-drug nanoplatform for stromal remodeling and enhanced chemotherapy via MMP3/TGF- β inhibition

doi: 10.1016/j.ijpx.2026.100489

Figure Lengend Snippet: In vitro therapeutic efficacy and synergy analysis of RPAE-QM in 4 T1 cells. (A) Cell viability of 4 T1 cells incubated with different formulations for 48 h determined by MTT assay. (B) The corresponding IC 50 values of the respective treatments. (C) Representative flow cytometry plots of Annexin V-FITC/PI staining for apoptosis analysis in 4 T1 cells. (D) Quantitative analysis of the total apoptotic rate. (E) Dose-response curves of free Que., free DM1, and their combination used for quantitative synergy determination. (F) The Combination Index (CI) plot as a function of Fraction affected (Fa) generated using the Chou-Talalay method; the reference line at CI = 1 indicates an additive effect, while CI < 1 indicates synergism. Data are presented as mean ± SD (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001 and **** P < 0.0001.

Article Snippet: After 24 h, the cells were harvested and stained with Annexin V-FITC/PI Cell Apoptosis Detection Kit (Beyotime, China) for flow cytometer analysis.

Techniques: In Vitro, Drug discovery, Incubation, MTT Assay, Flow Cytometry, Staining, Generated

Journal: bioRxiv

Article Title: Amplicon/Protein Bead Display enables quantitative in vitro biochemistry at scale

doi: 10.64898/2026.05.28.728566

Figure Lengend Snippet: a . Schematic of DNA templates encoding expression of a 25-variant FLAG library expressed as SNAP and eGFP fusion proteins on APBs. b . Schematic of experiments to detect binding between bead-displayed FLAG epitopes and M2 anti-FLAG antibodies using a Cy5-labeled secondary antibody. c . Representative brightfield and fluorescence images of APBs after incubation with M2 and Cy5-labeled secondary antibodies reporting on per-bead expression levels (GFP intensities), and antibody binding (Cy5 intensities). d . Box plots of per-bead median Cy5 pixel intensities (arbitrary units) for blank and GFP-positive beads. e . Schematic of single-bead sorting and sequencing workflow for validating genotype-phenotype linkages. FACS plot (far left) displays measured GFP (x-axis) and Cy5 (y-axis) intensities for the 384 GFP-positive beads sorted into a multi-well plate. f . Bar plots of expected (based on Poisson loading at λ = 0.1) and observed fractions of single-variant beads in the GFP-positive library. Error bars represent the Poisson-derived standard deviation. g . Measured Cy5 versus eGFP fluorescence intensities for all beads displaying three representative FLAG variants. Circles indicate single-variant beads; X’s indicate multi-variant beads. Red dashed lines indicate median Cy5/GFP ratio; gray shading shows interquartile range [0.25,0.75]. h . Median Cy5/GFP ratios calculated from all beads bearing a given variant versus single-variant beads only. Error bars denote interquartile range [0.25, 0.75]. i . Coefficient of variation (CV%) of median Cy5/GFP ratios across 1,000 bootstrap samples as a function of beads sampled per variant. j . Sequence logo showing relative position-normalized Cy5/GFP ratios for single amino acid substitutions (A, L, or E) at each position in the FLAG epitope. Red letters indicate wildtype residues; grey letters indicate substitutions.

Article Snippet: A fluorescence standard curve for estimating protein concentration on beads was generated using vortexed emulsion droplets containing known concentrations of recombinant eGFP protein (Origene).

Techniques: Expressing, Variant Assay, Binding Assay, Labeling, Fluorescence, Incubation, Sequencing, Derivative Assay, Standard Deviation, FLAG-tag