Journal: Bioactive Materials

Article Title: Spatiotemporally engineered microneedle for microenvironment remodeling propels mucosal regeneration after tracheal mucosal injury

doi: 10.1016/j.bioactmat.2026.01.026

Figure Lengend Snippet: Gel-AgNA/MgGA MN remodel the microenvironment after TMI by regulating infection, inflammation, oxidative stress, and vascular disruption. (a) FISH (marker of bacteria, red) staining of harvested samples after treated with Gel, Gel-AgNA, Gel-MgGA, and Gel-AgNA/MgGA MN. (b, c) Quantitative analysis of bacterial coverage and fluorescence intensity of FISH (n = 9). (d) Heatmaps of DEGs related to inflammation and oxidative stress. (e) RT-PCR analysis showing the relative expression levels of IL-1, TNF-α, HIF-1α, and IL-6 (n = 3). (f) IF staining of iNOS (marker of M1 macrophages, red) and CD206 (marker of M2 macrophages, green). (g) Quantitative analysis of iNOS and CD206 fluorescence intensity. (h) RT-PCR analysis showing the relative expression levels of Arg1 and CD206. (i) IF staining of CD31 (marker of blood vessels, red). (j, k) RT-PCR analysis showing the relative expression levels of VEGF and HIF-1α (n = 3).

Article Snippet: According to the method reported in previous study [ ], Fluorescence In Situ Hybridization (FISH) test was conducted for bacteria (Servicebio, Eub338) to assess bacterial distribution.

Techniques: Infection, Disruption, Marker, Bacteria, Staining, Fluorescence, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Kidney360

Article Title: The Positive Feedback Loop of Hypoxia-Inducible Factor-1 α /miR-295/Factor Inhibiting Hypoxia-Inducible Factor-1 in Hyperuricemic Nephropathy

doi: 10.34067/KID.0000001069

Figure Lengend Snippet: miR-295 is induced in renal tubules in HN mice. HN was induced by PO intraperitoneally and Ad orally administered for 2 weeks. Except for the control group, each group was intraperitoneally administered PO (350 mg/kg per day) and orally administered with Ad (70 mg/kg per day) to induce HN at 8:30 am for 14 consecutive days. Control mice were treated with normal saline. At the end of 21st day, the animals were euthanized. (A) Body weight changes in control and HN mice; (B) serum UA concentrations in each group; (C) serum CRE levels; (D) BUN levels; (E) representative images of HE staining, scale bar: 50 μ m; (F) representative images of Masson staining, scale bar: 50 μ m; (G) quantitative analysis of tubular injury score in different experimental groups; (H) quantification of Masson's trichrome–positive fibrotic area in kidney sections; (I) volcano plot of microRNA expression profiled by microarray in HN. Differentially expressed miRNAs were identified using data obtained from three biological replicates per experimental group ( n =3 mice per group, total six samples). Statistical significance was determined using the criteria of |log 2 fold change| >1.5 and P < 0.05. These thresholds were applied to ensure robust identification of meaningful expression differences. (J) qPCR analysis of miR-295 in mouse kidneys. The level of miR-295 was normalized to the level of U6 (internal loading control) of the same samples to determine the ratio with the ratio of control mice arbitrarily set as 1. (K) In situ hybridization showing miR-295 increase in kidney tissues. Representative images of double immunostaining with miR-295 and proximal tubule marker, LTL, scale bar: 50 μ m. (L) Quantitative analysis of fluorescence intensity. All the values are expressed as mean±SD ( n =6), * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Ad, adenine; CRE, creatinine; HE, hematoxylin–eosin; HN, hyperuricemic nephropathy; LTL, Lotus tetragonolobus lectin; PO, potassium oxonate; UA, uric acid

Article Snippet: The Digoxigenin-labeled mmu-miR-295 locked nucleic acid (LNA) probe and Fluorescence in situ hybridization (FISH) Kit were from Servicebio (Wuhan, China). miR-295 mimic, anti-miR-295 LNA, and FIH-1 siRNA were from Ruibo (Guangzhou, China).

Techniques: Control, Saline, Staining, Expressing, Microarray, In Situ Hybridization, Double Immunostaining, Marker, Fluorescence

Journal: bioRxiv

Article Title: Activity and retinoic acid drive hair cell spatial patterning in the zebrafish utricle

doi: 10.1101/2025.11.17.687290

Figure Lengend Snippet: A – C) Dorsal views of cabp1b and cabp2b HCR FISH in Tg(myo6b:GFP) at (A) 24, (B) 36, and (C) 48 hpf. Hair cells developed before 48 hpf are labeled by both probes (blue arrowheads). These specify into either striolar (yellow arrowheads) or extrastriolar (magenta arrowheads) hair cells during development. Scale bars = 20 µm. D) Hair cells are added in the first 10 days in both the utricle and saccule in an asynchronous and asymmetric manner. Utricle: n = 8 (24 hpf); 12 (30), 8 (36), 13 (48), 9 (72), 13 (120), 12 (240); Saccule: 8 (24), 9 (30), 7 (36), 13 (48), 9 (72), 3 (120), 2 (240). E) Relative proportions of hair cell subtypes change during larval development.

Article Snippet: Hybridization chain reaction (HCR) fluorescent in situ hybridization (FISH) (Molecular Instruments, HCR v3.0) was performed as directed for whole-mount zebrafish embryos and larvae ( , , ).

Techniques: Labeling

Journal: bioRxiv

Article Title: Activity and retinoic acid drive hair cell spatial patterning in the zebrafish utricle

doi: 10.1101/2025.11.17.687290

Figure Lengend Snippet: A – C) HCR FISH probing for cabp1b and cabp2b in Tg(myo6b:nlsEos) at 2, 5, and 10 dpf. First-developing hair cells were photoconverted (“PC”) at 36 hpf (dotted circle) to magenta while later-developing hair cells only have unconverted (cyan) Eos. Identity of all hair cells were tested with capb1b (“ES”) and capb2b . Scale bars = 20 µm. D-F) Polar plot summaries of 5 dpf utricles (B) (n = 11 utricles, 6 fish). PC hair cells (see triangles in E) are mostly located in extrastriola (F). Few PC hair cells were also intermediate (blue triangles in E). G) Summary of developmental data hair cell counts. H-I) Identity of PC hair cells by age (n = 11 ears, 8 fish (2 dpf); 10, 6 (5); 16, 8 (10)). By the end of larval development, most PC hair cells were still extrastriolar (ES), but a few were now striolar (white arrow heads in C)

Article Snippet: Hybridization chain reaction (HCR) fluorescent in situ hybridization (FISH) (Molecular Instruments, HCR v3.0) was performed as directed for whole-mount zebrafish embryos and larvae ( , , ).

Techniques:

Journal: bioRxiv

Article Title: Activity and retinoic acid drive hair cell spatial patterning in the zebrafish utricle

doi: 10.1101/2025.11.17.687290

Figure Lengend Snippet: A) HCR FISH probing for aldh1a3 (magenta) and cyp26b1 (yellow) shows complementary patterning of retinoic acid (RA) enzyme in 5 dpf Tg(myo6b:GFP) utricle. Scale bars = 20 µm. B) aldh1a3 (magenta) and cabp1b (yellow) indicates colocalization of RA synthesizing enzyme and extrastriolar hair cells. C) cabp2b (magenta) and cyp26b1 (yellow) indicates colocalization of RA degrading enzyme and striolar hair cells. D) At 2 dpf, aldh1a3 (magenta) and cyp26b1 (yellow) are polarized to either side of the developing maculae, where a few central hair cells contact neither end. E-F) 2 dpf hair cells expressing cabp1b contact aldh1a3 , as hair cells expressing cabp2b contact cyp26b1 .

Article Snippet: Hybridization chain reaction (HCR) fluorescent in situ hybridization (FISH) (Molecular Instruments, HCR v3.0) was performed as directed for whole-mount zebrafish embryos and larvae ( , , ).

Techniques: Expressing

Journal: bioRxiv

Article Title: Activity and retinoic acid drive hair cell spatial patterning in the zebrafish utricle

doi: 10.1101/2025.11.17.687290

Figure Lengend Snippet: A) Expression of cabp1b and cabp2b in Tg(myo6b:GFP); sputnik mutant fish at 5 dpf. White circles indicate many intermediate (double-labeled) hair cells present even at 5 dpf. B) HCR FISH after photoconversion at 36 hpf in Tg(myo6b:nlsEos) at 5 dpf. White circle indicates hair cell that is not photoconverted but still double-labeled. Scale bar = 20 µm. C-D) Sputnik mutants (“Sput”) (n = 21 ears, 15 fish) have fewer hair cells and significantly different proportions of striolar/extrastriolar/intermediate hair cells at 5 dpf relative to wildtype (“WT”) fish (n = 12, 8). E-F) Hair cells photoconverted at 36 hpf in sputnik fish with Tg(myo6:nlsEos) (n = 7, 7) show that most early-developing hair cells stay in the intermediate state (vs wildtype, data from ). However, there are fewer photoconverted hair cells than double-labeled hair cells, indicating some later-developing hair cells are intermediate. G) Polar plot summary of 5 dpf utricles from G) Tg(myo6b:GFP) (n =12, 9). Intermediate hair cells were located along the zonal boundary. H) Polar plot summary of Tg(myo6b:nlsEos) (n = 7, 6). PC hair cells (triangles) are mostly located in extrastriola. Most PC hair cells were also intermediate (blue triangles) and located along the zonal boundary.

Article Snippet: Hybridization chain reaction (HCR) fluorescent in situ hybridization (FISH) (Molecular Instruments, HCR v3.0) was performed as directed for whole-mount zebrafish embryos and larvae ( , , ).

Techniques: Expressing, Mutagenesis, Labeling