polymyxin resistant mutant mut s  (ATCC)

Journal: Nucleic Acids Research

Article Title: PhoP-regulated VirK acts as an accessory factor to maintain virulence in polymyxin-resistant Klebsiella pneumoniae

doi: 10.1093/nar/gkag290

Figure Lengend Snippet: Transcriptomic analysis of Mut-S in comparison to Kpn2146 strains. ( A ) Volcano map displaying the upregulated (red dots) and downregulated (yellow dots) genes between the Mut-S group and the control group. ( B ) Changes in the relative expression of the mgrB, phoP , and virK genes in the Mut-S strain compared with those in the WT strain. ( C )Changes in the virK gene expression in the Mut-S strain after supplementation with the mgrB gene. ( D ) Changes in phoP and virK gene expressions after phoP gene knockdown in the Mut-S strain. ( E ) Changes in the mgrB and virK gene expressions after the mgrB gene was complemented in the phoP knockout strain in Mut-S. ( F ) Changes in the phoP and virK gene expressions after the phoP gene was supplemented in the phoP knockout strains in Mut-S. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: During preliminary investigations of a polymyxin-resistant mutant (Mut-S) of K. pneumoniae ATCC BAA-2146 (Kpn2146) [ ], we detected highly upregulated expression of the Kpn2146:RS17285 gene.

Techniques: Comparison, Control, Expressing, Gene Expression, Knockdown, Knock-Out

Journal: bioRxiv

Article Title: Hyperactive STAT1 Promotes T Follicular Helper Type 1 Cell Differentiation to Trigger Autoimmunity

doi: 10.64898/2026.03.19.713058

Figure Lengend Snippet: ( A ) Sixteen pathogenic mutations in STAT1 coding region carried by 20 patients are indicated. Red text denotes patients with autoimmune phenotypes. The asterisks (*) indicate newly identified patients. ( B ) Heatmap illustrating autoimmune phenotypes in patients. Red and grey shading represents positive and negative status, respectively. ( C ) Representative flow cytometry plots and pooled data of the percentage of circulating CXCR5 + T cells in CD3 + CD4 + CD45RO + T cells. ( D ) Representative flow cytometry plots and pooled data of the percentage of circulating CXCR5 + PD-1 + in CD3 + CD4 + CD45RO + T cells. ( E ) Bar plots showing PD-1 MFI (normalized to aged-matched controls). ( F ) Representative flow cytometry plots and pooled data of the percentage of cTfh subsets in circulating CXCR5 + CD45RA - CD4 + T cells. MFI: Mean fluorescence intensity. cTfh: circulating Tfh. Numbers adjacent to outlined areas or in the indicated quadrants represent the percentage of cells in the area. Each symbol represents an individual throughout. Data were represented as means ± SE and analyzed with two-tailed unpaired t test.

Article Snippet: Mutant STAT1 (p.T385M) mice carrying a missense mutation (c.1154C>T) were generated by in situ gene targeting using TurboKnockout ® technology (Cyagen Biosciences, China).

Techniques: Flow Cytometry, Fluorescence, Two Tailed Test

Journal: bioRxiv

Article Title: Hyperactive STAT1 Promotes T Follicular Helper Type 1 Cell Differentiation to Trigger Autoimmunity

doi: 10.64898/2026.03.19.713058

Figure Lengend Snippet: ( A ) Histogram of pSTAT1 (Y701) and total STAT1 in B cells and CD4 + T cells. ( B ) pSTAT1 and total STAT1 MFI in indicated subsets. ( C ) H&E staining of liver sections (left) and lung sections (right). ( D ) Bar graph showing numbers of mice with positive or negative inflammatory cell infiltration in indicated tissue. ( E-F ) The levels of anti-dsDNA antibody detected in serum in mice at 6-8 weeks (E) and 20-22 (F) weeks. ( G ) Representative Hep-2 ANA (up) and Crithidia luciliae anti-dsDNA antibody (down) staining. ( H ) Bar graph showing numbers of mice with positive or negative results in indicated autoantibody. ( I ) Representative staining of IgG in kidney. ( J ) Bar graph showing numbers of mice with positive or negative IgG deposition in kidney. Each symbol represents an individual mouse throughout. Data are representative of three independent experiments. Data were represented as means ± SEM and analyzed with two-tailed unpaired t test.

Article Snippet: Mutant STAT1 (p.T385M) mice carrying a missense mutation (c.1154C>T) were generated by in situ gene targeting using TurboKnockout ® technology (Cyagen Biosciences, China).

Techniques: Staining, Two Tailed Test

Journal: bioRxiv

Article Title: Hyperactive STAT1 Promotes T Follicular Helper Type 1 Cell Differentiation to Trigger Autoimmunity

doi: 10.64898/2026.03.19.713058

Figure Lengend Snippet: (A-C) The indicated mice were intraperitoneally immunized with KLH/CFA. 14 days after the immunization, splenic Tfh cells were determined. Representative flow cytometry plots and pooled data of the percentage and numbers of splenic CXCR5 + PD-1 + and CXCR5 + Bcl-6 + Tfh cells in CD4 + CD44 + Foxp3 - T cells. (D-E) The indicated mice were intraperitoneally immunized with NP-KLH/CFA. 14 days after the immunization, the percentages and numbers of splenic CXCR5 + PD-1 + and CXCR5 + Bcl-6 + Tfh cells were determined. (F-G) The indicated mice were immunized subcutaneously with KLH/CFA. The percentages of Tfh at indicated time points were determined. (H-M) Naïve OT-II cells from indicated mice were isolated and transferred into congenic mice. The recipient mice were subcutaneously immunized with OVA/CFA. Seven days after the immunization, Tfh cells in dLNs were determined. (H) Experimental design for evaluating the role of mutant Stat1 in Tfh differentiation. (I-J) Representative flow cytometry plots and pooled data of the percentage and number of donor cells. (K-L) Representative flow cytometry plots and pooled data of the percentage of CXCR5 + PD-1 + and CXCR5 + Bcl-6 + in donor cells. (M) MFI of CXCR5, PD-1 and Bcl-6 in donor cells. Numbers adjacent to outlined areas represent the percentage of cells in the area. Each symbol represents an individual throughout. Data are representative of three independent experiments. Data were represented as means ± SEM and analyzed with two-tailed unpaired t test.

Article Snippet: Mutant STAT1 (p.T385M) mice carrying a missense mutation (c.1154C>T) were generated by in situ gene targeting using TurboKnockout ® technology (Cyagen Biosciences, China).

Techniques: Flow Cytometry, Isolation, Mutagenesis, Two Tailed Test

Journal: bioRxiv

Article Title: Hyperactive STAT1 Promotes T Follicular Helper Type 1 Cell Differentiation to Trigger Autoimmunity

doi: 10.64898/2026.03.19.713058

Figure Lengend Snippet: ( A-B) Volcano plots showing DEGs of non-Tfh (A) and Tfh (B) between WT and Stat1 Mut/WT . Cells were sorted from mice immunized with KLH/CFA for 14 days. (C-D) GSEA plot for the indicated gene set. (E) Heatmap visualization of DEGs in non-Tfh and Tfh between WT and Stat1 Mut/WT mice immunized with KLH/CFA for 14 days. (F) PCA plot of the transcriptomic data of indicated cell groups from mice at age of 20-22 weeks. Each dot represents an individual sample. (G) Heatmap visualization of DEGs in non-Tfh and Tfh between WT and Stat1 Mut/WT at age of 20-22 weeks. DEGs: differentially expressed genes. GSEA: gene set enrichment analysis. PCA: principal component analysis.

Article Snippet: Mutant STAT1 (p.T385M) mice carrying a missense mutation (c.1154C>T) were generated by in situ gene targeting using TurboKnockout ® technology (Cyagen Biosciences, China).

Techniques:

Journal: bioRxiv

Article Title: Hyperactive STAT1 Promotes T Follicular Helper Type 1 Cell Differentiation to Trigger Autoimmunity

doi: 10.64898/2026.03.19.713058

Figure Lengend Snippet: ( A ) Heatmap for genome-wide distribution of STAT1 and mutant STAT1 peaks at TSS in non-Tfh ang Tfh cells. ( B ) The STAT1 and mutant STAT1 binding site is identical to the GAS motif. ( C-E ) Venn diagrams showing the genes regulated by STAT1 (mutant STAT1) and STAT1(mutant STAT1)-bound genes between different cells as indicated. Differentially expressed genes were marked with purple. Differentially bound genes were marked with red. ( F ) Tracks of STAT1 (mutant STAT1)-binding peaks located at gene loci including Stat1 , Ifng , Tbx21 , Cxcr5 , Pdcd1 and Bcl6 . ( G ) Heatmap for DEGs of CD4 + naïve T cells between WT and Stat1 Mut/WT mice. ( H ) CD4 + CD25 - CD44 - CD62L + naïve CD4 + T cells isolated from WT and Stat1 Mut/WT mice, and the mRNA expression level of indicated genes was determined by quantitative PCR. Data were represented as means ± SEM and analyzed with two-tailed unpaired t test.

Article Snippet: Mutant STAT1 (p.T385M) mice carrying a missense mutation (c.1154C>T) were generated by in situ gene targeting using TurboKnockout ® technology (Cyagen Biosciences, China).

Techniques: Genome Wide, Mutagenesis, Binding Assay, Isolation, Expressing, Real-time Polymerase Chain Reaction, Two Tailed Test

Journal: bioRxiv

Article Title: Hyperactive STAT1 Promotes T Follicular Helper Type 1 Cell Differentiation to Trigger Autoimmunity

doi: 10.64898/2026.03.19.713058

Figure Lengend Snippet: ( A ) Representative flow cytometry plots and pooled data of the percentage of IFN-γ + and IL-17A + in CD3 + CD4 + CXCR5 + PD-1 + cTfh cells from STAT1 -GOF patients and controls. ( B ) MFI of T-bet in cTfh cells from STAT1 -GOF patients and controls. ( C ) Representative flow cytometry plots and pooled data of the percentage of IFN-γ + in splenic Tfh of mice at age of 20-22 weeks. ( D ) Percentages of IFN-γ + in Tfh of mice after immunization with KLH/CFA for indicated time points. ( E ) Percentage of IFN-γ + in Tfh from indicated donor cells. ( F ) Isolated Tfh from indicated mice and B cell from WT co-cultured in vitro . IFN-γ level in supernatant, percentage of GC B cells and IFNγR, T-bet MFI in GC B cells were determined. ( G ) T-bet MFI in splenic GC B cells of mice at age of 6-8 weeks. ( H ) MFI of pSTAT1 (Y701) and total STAT1 in B cells and CD4 + T cells from mice injected with anti-IFN-γ antibody or isotype control. ( I ) Percentage of splenic GC B cells and plasma B cells in indicated mice. ( J ) MFI of T-bet in splenic GC B cells in indicated mice. ( K ) Percentage of IgG2a + in GC B cells in indicated mice. ( L ) Serum anti-dsDNA antibody level in indicated mice was determined. Each symbol represents an individual throughout. Data were represented as means ± SEM and analyzed with two-tailed paired(A) or unpaired t test(B-L).

Article Snippet: Mutant STAT1 (p.T385M) mice carrying a missense mutation (c.1154C>T) were generated by in situ gene targeting using TurboKnockout ® technology (Cyagen Biosciences, China).

Techniques: Flow Cytometry, Isolation, Cell Culture, In Vitro, Injection, Control, Clinical Proteomics, Two Tailed Test