Journal: Frontiers in Microbiology

Article Title: Genomic characterization of a pathogenic Bacillus licheniformis strain LSDY01: deciphering its genetic diversity and virulence-associated traits

doi: 10.3389/fmicb.2026.1815181

Figure Lengend Snippet: Genome visualization and annotation of strain LSDY01. (A) Chromosome; (B) plasmid. The element colour of each circle is indicated in the legend; (C) average Nucleotide Identity (ANI) heatmap illustrating genomic similarity among selected Bacillus strains. LSDY01 shows the highest ANI (99.6%) with Bacillus licheniformis ATCC 14580 and DSM 13, confirming its taxonomic assignment to this species. The color scale represents percentage similarity.

Article Snippet: For comparative genomic analysis with Bacillus licheniformis , a total of 498 B. licheniformis genome sequences (October 2025) were retrieved from the National Center for Biotechnology Information (NCBI).

Techniques: Plasmid Preparation

Journal: Frontiers in Microbiology

Article Title: Genomic characterization of a pathogenic Bacillus licheniformis strain LSDY01: deciphering its genetic diversity and virulence-associated traits

doi: 10.3389/fmicb.2026.1815181

Figure Lengend Snippet: Molecular typing and genomic phylogeny of Bacillus licheniformis strain LSDY01. (A) In silico multi-locus sequence typing (MLST) reveals that LSDY01 belongs to the uncommon ST20, differing from the prevalent ST3 by a single allele. (B) cgMLST Phylogenetic analysis places LSDY01 in a clade with Daqu-derived strains CP143961.1 and CP143962.1 (bootstrap = 100), indicating recent divergence.

Article Snippet: For comparative genomic analysis with Bacillus licheniformis , a total of 498 B. licheniformis genome sequences (October 2025) were retrieved from the National Center for Biotechnology Information (NCBI).

Techniques: In Silico, Sequencing, Derivative Assay

Journal: Frontiers in Microbiology

Article Title: Genomic characterization of a pathogenic Bacillus licheniformis strain LSDY01: deciphering its genetic diversity and virulence-associated traits

doi: 10.3389/fmicb.2026.1815181

Figure Lengend Snippet: BRIG-based comparative genomic analysis of Bacillus licheniformis LSDY01. (A) Circular comparison of the LSDY01 chromosome with closely related strains CP143961.1 and CP143962.1 . A unique ~157 kb genomic island (position ~2.3–2.5 Mb) was identified, characterized by significantly lower GC content (33.03%) and the co-localization of an anti-CRISPR gene (AcrIIA7) and a yoqS locus. (B) Plasmid pLSDY01 alignment with reference sequences CP076292.1 and CP178437.1 , showing high identity (97.66%) and coverage (99%). The Pistol gene is uniquely present in pLSDY01. (C) Linear comparison of the ~157 kb genomic island with those of CP154901.1 and CP143961.1 . The gray shading indicates regions of shared homology among different elements. The open reading frames are marked by colored arrows.

Article Snippet: For comparative genomic analysis with Bacillus licheniformis , a total of 498 B. licheniformis genome sequences (October 2025) were retrieved from the National Center for Biotechnology Information (NCBI).

Techniques: Comparison, CRISPR, Plasmid Preparation

Journal: Frontiers in Microbiology

Article Title: Genomic characterization of a pathogenic Bacillus licheniformis strain LSDY01: deciphering its genetic diversity and virulence-associated traits

doi: 10.3389/fmicb.2026.1815181

Figure Lengend Snippet: Phylogenetic distribution of virulence-associated genes in Bacillus licheniformis strains. Maximum-likelihood tree of 336 Bacillus licheniformis genomes. Notably, yoqS, yoqJ , and Pistol were uniquely identified in strain LSDY01, while hrtA was present only in a subset of strains, highlighting the distinct gene profile of LSDY01 compared to other Bacillus licheniformis isolates.

Article Snippet: For comparative genomic analysis with Bacillus licheniformis , a total of 498 B. licheniformis genome sequences (October 2025) were retrieved from the National Center for Biotechnology Information (NCBI).

Techniques:

Journal: Frontiers in Microbiology

Article Title: Genomic characterization of a pathogenic Bacillus licheniformis strain LSDY01: deciphering its genetic diversity and virulence-associated traits

doi: 10.3389/fmicb.2026.1815181

Figure Lengend Snippet: Strong biofilm formation by Bacillus licheniformis LSDY01. Biofilm biomass (OD 570 ) was measured by crystal violet staining after 36 h at 37 °C in TSB with 1% glucose. LSDY01 (0.67 ± 0.03) was classified as a strong producer. Error bars: SD of quadruplicate wells.

Article Snippet: For comparative genomic analysis with Bacillus licheniformis , a total of 498 B. licheniformis genome sequences (October 2025) were retrieved from the National Center for Biotechnology Information (NCBI).

Techniques: Staining

Journal: Frontiers in Microbiology

Article Title: Genomic characterization of a pathogenic Bacillus licheniformis strain LSDY01: deciphering its genetic diversity and virulence-associated traits

doi: 10.3389/fmicb.2026.1815181

Figure Lengend Snippet: Effect of Bacillus licheniformis LSDY01 on HEK293 cell viability after 24 h co-culture. CCK-8 was added and absorbance (OD₄₅₀) was measured at the indicated time points. All values were normalized by subtracting the background signal of cell-free medium. Data are presented as mean ± SD ( n = 3). Statistical significance was determined by independent samples t -test (two-tailed). At the 2 h, 4 h and 6 h readings, no significant differences were observed between the bacterial co-culture group and the control group ( p > 0.05 for all).

Article Snippet: For comparative genomic analysis with Bacillus licheniformis , a total of 498 B. licheniformis genome sequences (October 2025) were retrieved from the National Center for Biotechnology Information (NCBI).

Techniques: Co-Culture Assay, CCK-8 Assay, Two Tailed Test, Control

Journal: EMBO Molecular Medicine

Article Title: A new form of diabetes caused by INS mutations defined by zygosity, stem cell and population data

doi: 10.1038/s44321-025-00362-9

Figure Lengend Snippet: Unrelated control (black), heterozygous R6C (HET, yellow) and isogenic corrected (HET CORR, blue) iPSCs were differentiated into β cells. ( A ) iPSC-islet lysates were analyzed by SDS-PAGE under reducing conditions, electrotransfer to nitrocellulose, and immunoblotting with anti-human proinsulin. The blot was cropped and rearranged for clarity. The left panel is reused in Fig. as it represents a shared control. ( B ) Quantification of preproinsulin to proinsulin ratio from ( A ) (unrelated control n = 3, HET CORR n = 3, HET n = 4). ( C ) iPSC-islet lysates were resolved by nonreducing 12%-NuPAGE, and the completed gel was then treated with 100 mM DTT at 60 °C for 10 min before electrotransfer to nitrocellulose and immunoblotting with anti-proinsulin. n = 2. Medium from Min6 β cells transfected with human proinsulin was used as a positive control (INS, lane next to marker, M). The blot was cropped and rearranged for clarity. The left panel is reused in Fig. as it represents the shared marker and positive control. ( D ) Proinsulin and ( E ) insulin content (ng) normalized to total protein content (μg). ( F ) Proinsulin to insulin content ratio from ( D , E ). ( D – F ) HET CORR n = 4, HET n = 7. All panels: Unpaired t -test. In box plots, the median of independent experiments is shown by a horizontal line; 25 th and 75 th percentiles are at the bottom and top of the boxes; whiskers represent the minimum and maximum values. .

Article Snippet: Mouse anti-human proinsulin B-C junction sequence KTRREAEDLQ , Abmart , Cat# B-C junction; RRID: AB_2921300.

Techniques: Control, SDS Page, Electrotransfer, Western Blot, Transfection, Positive Control, Marker

Journal: EMBO Molecular Medicine

Article Title: A new form of diabetes caused by INS mutations defined by zygosity, stem cell and population data

doi: 10.1038/s44321-025-00362-9

Figure Lengend Snippet: Unrelated control (black), homozygous R6C (HOM, pink) and isogenic corrected (HOM CORR, blue) iPSCs were differentiated into β cells. ( A ) iPSC-islet lysates were analyzed by SDS-PAGE under reducing conditions, electrotransferred to nitrocellulose, and immunoblotted with anti-human proinsulin. The blot was cropped and rearranged for clarity. The left panel is reused as in Fig. , as it represents a shared control. ( B ) Quantification of preproinsulin to proinsulin ratio from ( A ) (unrelated control n = 3, HOM CORR n = 3, HOM n = 8) and ( C ) of homozygous R6C iPSC-β cells treated with vehicle DMSO or MG132 (10 μM) for 30 min ( n = 4). ( D ) iPSC-islet lysates were resolved by nonreducing 12%-NuPAGE and the completed gel then treated with 100 mM DTT at 60 °C for 10 min before electrotransfer to nitrocellulose and immunoblotting with anti-proinsulin. n = 2. Medium from Min6 β cells transfected with human proinsulin was used as a positive control (INS, lane next to marker, M). The blot was cropped and rearranged for clarity. The left panel is reused as in Fig. as it represents a shared marker and a positive control. ( E ) Proinsulin and ( F ) insulin content normalized to total protein content. ( G ) Proinsulin to insulin content ratio from ( E , F ). ( E – G ) HOM CORR n = 8, HOM n = 12–13. ( H ) Insulin content normalized to total protein content along stage 7 (S7, HOM CORR n = 5, HOM n = 12), 1 week (LC1W, HOM CORR n = 3, HOM n = 3), 2 weeks (LC2W, HOM CORR n = 3, HOM n = 3), 3 weeks (LC3W, HOM CORR n = 3, HOM n = 3), and 4 weeks (LC4W) of long culture (LC, HOM CORR n = 12, HOM n = 15). All panels: Unpaired t -test. In box plots, the median of independent experiments is shown by a horizontal line; 25 th and 75 th percentiles are at the bottom and top of the boxes; whiskers represent the minimum and maximum values. .

Article Snippet: Mouse anti-human proinsulin B-C junction sequence KTRREAEDLQ , Abmart , Cat# B-C junction; RRID: AB_2921300.

Techniques: Control, SDS Page, Electrotransfer, Western Blot, Transfection, Positive Control, Marker

Journal: EMBO Molecular Medicine

Article Title: A new form of diabetes caused by INS mutations defined by zygosity, stem cell and population data

doi: 10.1038/s44321-025-00362-9

Figure Lengend Snippet: Heterozygous R6C (HET, yellow) and isogenic corrected (HET CORR, blue) iPSCs were differentiated into long-cultured β cells. ( A – C ) Static proinsulin and insulin secretion in response to 2.8 mM glucose (G2.8), 16.8 mM glucose (G16.8), or 16.8 mM glucose plus 10 μM forskolin (G16.8 + Fk). HET CORR n = 4, HET n = 7. ( A ) Proinsulin and ( B ) insulin secretion (ng) normalized to protein content (μg). ( C ) Proinsulin to insulin ratio from ( A , B ). ( D – H ) Dynamic insulin secretion upon perifusion with 2.8 mM glucose, 16.8 mM glucose (G16.8), G16.8 plus exendin-4 (Ex4, 50 ng/mL, 11.8 nM) or G2.8 plus KCl (30 mM). HET CORR n = 3, HET n = 4. ( D ) Insulin secretion normalized to protein content, with ( E – H ) area under the curve (AUC) per minute of secretion at G2.8, G16.8, G16.8 + Ex4, and G2.8 + KCl. All panels: Unpaired t -test. In box plots, the median of independent experiments is shown by a horizontal line; 25 th and 75 th percentiles are at the bottom and top of the boxes; whiskers represent the minimum and maximum values. In time course line plots, data are shown as mean ± s.e.m. .

Article Snippet: Mouse anti-human proinsulin B-C junction sequence KTRREAEDLQ , Abmart , Cat# B-C junction; RRID: AB_2921300.

Techniques: Cell Culture

Journal: EMBO Molecular Medicine

Article Title: A new form of diabetes caused by INS mutations defined by zygosity, stem cell and population data

doi: 10.1038/s44321-025-00362-9

Figure Lengend Snippet: Homozygous R6C (HOM, pink) and isogenic corrected (HOM CORR, blue) iPSCs were differentiated into long-cultured β cells. ( A – C ) Static proinsulin and insulin secretion in response to 2.8 mM glucose (G2.8), 16.8 mM glucose (G16.8), or 16.8 mM glucose plus 10 μM forskolin (G16.8 + Fk). HOM CORR n = 8, HOM n = 12 (for proinsulin), HOM n = 13 (for insulin). ( A ) Proinsulin and ( B ) insulin secretion normalized to protein content. ( C ) Proinsulin to insulin ratio from ( A , B ). ( D – I ) Dynamic insulin secretion upon perifusion with 2.8 mM glucose, 16.8 mM glucose (G16.8), G16.8 plus exendin-4 (Ex4, 50 ng/mL, 11.8 nM), or G2.8 plus KCl (30 mM). HOM CORR n = 12, HOM n = 14. ( D ) Insulin secretion normalized to protein content, with ( E ) zoom in on 16.8 mM glucose response. ( F – I ) Area under the curve (AUC) per minute of secretion at G2.8, G16.8, G16.8 + Ex4 and G2.8 + KCl. All panels: Unpaired t -test. In box plots, the median of independent experiments is shown by a horizontal line; 25 th and 75 th percentiles are at the bottom and top of the boxes; whiskers represent the minimum and maximum values. In time course line plots, data are shown as mean ± s.e.m. .

Article Snippet: Mouse anti-human proinsulin B-C junction sequence KTRREAEDLQ , Abmart , Cat# B-C junction; RRID: AB_2921300.

Techniques: Cell Culture