Journal: bioRxiv

Article Title: OmpA controls intracellular survival of Acinetobacter baumannii through TFEB activation and lysosomal remodeling

doi: 10.64898/2026.04.18.719357

Figure Lengend Snippet: (A) Immunostaining of iPLA 2 in HeLa cells infected with the ATCC 17978 strain for 24 h. (B) Immunoblot analysis of inactive and active forms of iPLA 2 in HeLa cells infected with the ATCC 17978 strain for 24 h. (C) Enzymatic activity of iPLA 2 in HeLa cells infected with the ATCC 17978 strain for 24 h, in the presence or absence of BEL. * P =0.025 for ATCC 17978 vs. CTL, ** P =0.05 for ATCC 17978 vs. ATCC 1797 8 + BEL (5 µM), *** P =0.042 for ATCC 17978 vs. ATCC 17978 + BEL (10 µM). (D) Cox-2 enzymatic activity in HeLa cells infected with the ATCC 17978 strain for 24 h, in the presence or absence of BEL and NS. (* P =0.0011 for ATCC 17978 vs. CTL, * P =0.049 for CTL vs. ATCC 17978 + BEL (5 µM), * P =0.0096 for CTL vs. ATCC 17978 + NS, ** P =0.0005 for ATCC 17978 vs. ATCC 17978 + BEL (5 µM), *** P =0.001 for ATCC 17978 vs. ATCC 17978 + NS. ( E ) PGE2 production in HeLa cells infected with the ATCC 17978 strain for 24 h, in the presence or absence of BEL and NS. * P =0.004 for ATCC 17978 vs. CTL, ** P =0.0417 for ATCC 17978 vs. ATCC 17978 + BEL (5 µM), *** P =0.0015 for ATCC 17978 vs. ATCC 17978 + NS. CTL: control; Str: staurosporine; BEL: bromoelone lactone; NS: cyclooxygenase inhibitor; Cox-2: cyclooxygenase-2; PGE2: prostaglandin 2.

Article Snippet: Briefly, HeLa cells plated on coverslips were incubated with A. baumannii ATCC 17978 (1 x 10 7 CFU/mL) or stauroporine (10 mg/L, apopotosis inducer) for 24 h at 37 °C, 5% CO 2 .

Techniques: Immunostaining, Infection, Western Blot, Activity Assay, Control

Journal: bioRxiv

Article Title: OmpA controls intracellular survival of Acinetobacter baumannii through TFEB activation and lysosomal remodeling

doi: 10.64898/2026.04.18.719357

Figure Lengend Snippet: (A) Immnunoblot analysis of TFEB in HeLa cells infected with the ATCC 17978 strain for 2 h in the presence or absence of BEL (5 µM). * P =0.041 for ATCC 17978 vs. ATCC 17978 + BEL. (B) TFEB nuclear translocation in TFEB GFP+ cells infected with the ATCC 17978 strain for 2 h in the presence or absence of BEL (5 µM). * P =0.0162 for CTL vs. ATCC 17978, * P =0.0452 for CTL vs. ATCC 17978, ** P =0.0497 for ATCC 17978 vs. ATCC 17978 + BEL. (C) Immunoblot analysis of TFEB in HeLa cells transfected with scrambled or iPLA 2 siRNA for 48 h and infected with the ATCC 17978 strain for 2 h. * P =0.0012 for CTL vs. siRNA of iPLA 2 . BEL: bromoelone lactone; Sc: scramble.

Article Snippet: Briefly, HeLa cells plated on coverslips were incubated with A. baumannii ATCC 17978 (1 x 10 7 CFU/mL) or stauroporine (10 mg/L, apopotosis inducer) for 24 h at 37 °C, 5% CO 2 .

Techniques: Infection, Translocation Assay, Western Blot, Transfection

Journal: bioRxiv

Article Title: OmpA controls intracellular survival of Acinetobacter baumannii through TFEB activation and lysosomal remodeling

doi: 10.64898/2026.04.18.719357

Figure Lengend Snippet: (A) LPC production in HeLa cells infected with the ATCC 17978 strain for 24 h, in the presence or absence of BEL. * P <0.0001 for ATCC 17978 vs. CTL, * P =0.032 for CTL vs. ATCC 17978 + BEL, ** P =0.048 for ATCC 17978 vs. ATCC 17978 + BEL. (B) TFEB activation in TFEB GFP+ cells treated with LPC (0.5 µM) for 1 h. * P <0.0001 for 0 h vs. 1 h. CTL: control; BEL: bromoelone lactone; LPC: lysophosphatidylcholine.

Article Snippet: Briefly, HeLa cells plated on coverslips were incubated with A. baumannii ATCC 17978 (1 x 10 7 CFU/mL) or stauroporine (10 mg/L, apopotosis inducer) for 24 h at 37 °C, 5% CO 2 .

Techniques: Infection, Activation Assay, Control

Journal: bioRxiv

Article Title: OmpA controls intracellular survival of Acinetobacter baumannii through TFEB activation and lysosomal remodeling

doi: 10.64898/2026.04.18.719357

Figure Lengend Snippet: (A) Inhibition of calcium entry through ORAI channel reduces TFEB activation in TFEB GFP+ cells infected with the ATCC 17978 strain for 2 h. * P =0.05 for ATCC 17978 vs. ATCC 17978 + GSK. 7975A (B) Chelation of intracellular calcium with EGTA reduces TFEB activation in TFEB GFP+ cells infected with the ATCC 17978 strain for 2 h. * P <0.0001 for CTL vs. ATCC 17978, * P <0.0001 for CTL vs ATCC 17978 + EGTA, ** P <0.0001 for ATCC 17978 vs. ATCC 17978 + EGTA. (C) Immunoblot analysis of ORAI and TFEB in HeLa cells transfected with scrambled or iPLA 2 siRNA for 48 h and infected with the ATCC 17978 strain for 2 h. * P =0.0093 for ATCC 17978 vs. siRNA ORAI. (D) Lysosomal analysis in HeLa cells infected with the ATCC 17978 strain for 2 h. Acidic organelles were labeled with LysoTracker Red and mitochondria were labeled with MitoTracker Green. CTL: control, EGTA: Ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid tetrasodium salt.

Article Snippet: Briefly, HeLa cells plated on coverslips were incubated with A. baumannii ATCC 17978 (1 x 10 7 CFU/mL) or stauroporine (10 mg/L, apopotosis inducer) for 24 h at 37 °C, 5% CO 2 .

Techniques: Inhibition, Activation Assay, Infection, Western Blot, Transfection, Labeling, Control

Journal: bioRxiv

Article Title: OmpA controls intracellular survival of Acinetobacter baumannii through TFEB activation and lysosomal remodeling

doi: 10.64898/2026.04.18.719357

Figure Lengend Snippet: (A) Ammonia production in HeLa cells infected with the ATCC 17978 strain for 2 h. * P =0.011 for ATCC 17978 vs. CTL. (B) Intracellular replication of the ATCC 17978 strain in HeLa cells infected at 2, 4 and 8 h post-infection, in the presence or absence of bafilomycin. Data are presented as fold change relative to the intracellular CFU at 2 h. * P =0.033 for ATCC 17978 vs. ATCC 17978 + bafilomycin. (C) Lysosomal pH measurement of HeLa cells infected for 2 h with the ATCC 17978 strain stained with LysoSensor Yellow/Blue. * P =0.0164 for ATCC 17978 vs. CTL. (D) pH measurement of LB medium over 24 h in the presence of the ATCC 17978 strain. (E) A. baumannii ATCC 17978 in LB medium during 24 h under acidic or neutral conditions. CTL: control.

Article Snippet: Briefly, HeLa cells plated on coverslips were incubated with A. baumannii ATCC 17978 (1 x 10 7 CFU/mL) or stauroporine (10 mg/L, apopotosis inducer) for 24 h at 37 °C, 5% CO 2 .

Techniques: Infection, Staining, Control

Journal: bioRxiv

Article Title: OmpA controls intracellular survival of Acinetobacter baumannii through TFEB activation and lysosomal remodeling

doi: 10.64898/2026.04.18.719357

Figure Lengend Snippet: (A) Adherence and invasion of ATCC 17978 and ATCC 17978 ΔOmpA strains in HeLa cells for 2 h. * P =0.003 for ATCC 17978 vs. ATCC 17978 ΔOmpA (adherence), * P =0.032 for ATCC 17978 vs. ATCC 17978 ΔOmpA (invasion). (B) Enzymatic activity of iPLA 2 in HeLa cells infected with ATCC 17978 or ATCC 17978 ΔOmpA strains for 24 h. * P =0.025 for CTL vs. ATCC 17978, * P =0.0255 for CTL vs. ATCC 17978 ΔOmpA. (C) LPC production in HeLa cells infected with ATCC 17978 or ATCC 17978 ΔOmpA strains for 24 h. * P <0.001 for CTL vs. ATCC 17978, * P =0.043 for CTL vs. ATCC 17978 ΔOmpA, ** P =0.03 for ATCC 17978 vs. ATCC 17978 ΔOmpA. (D) Immunoblot analysis of TFEB in HeLa cells infected with ATCC 17978 or ATCC 17978 ΔOmpA strains for 2 h. * P =0.0424 for CTL vs. ATCC 17978. CTL: control.

Article Snippet: Briefly, HeLa cells plated on coverslips were incubated with A. baumannii ATCC 17978 (1 x 10 7 CFU/mL) or stauroporine (10 mg/L, apopotosis inducer) for 24 h at 37 °C, 5% CO 2 .

Techniques: Activity Assay, Infection, Western Blot, Control

Journal: bioRxiv

Article Title: OmpA controls intracellular survival of Acinetobacter baumannii through TFEB activation and lysosomal remodeling

doi: 10.64898/2026.04.18.719357

Figure Lengend Snippet: (A and B) Lysosomal analysis in HeLa cells infected with ATCC 17978 or ATCC 17978 ΔOmpA strains for 2 h. Acidic organelles were labeled with LysoTracker Red and mitochondria were labeled with MitoTracker Green. Lysosomal staining in infected HeLa cells is presented in the bar graph as a percentage relative to non-infected control cells. * P =0.0111 for ATCC 17978 vs CTL. ** P =0.0140 for ATCC 17978 vs. ATCC 17978 ΔOmpA. (C) Ammonia production in HeLa cells infected with ATCC 17978 or ATCC 17978 ΔOmpA strains for 2 h.* P =0.011 for ATCC 17978 vs CTL, ** P =0.03031 for ATCC 17978 vs ATCC 17978 ΔOmpA. (D) Intracellular replication of ATCC 17978 or ATCC 17978 ΔOmpA strains in HeLa cells infected at 2, 4 and 8 h post-infection, in the presence or absence of bafilomycin. Data are expressed as fold change in intracellular CFU relative to 2 h.* P =0.033 for ATCC 17978 vs. ATCC 17978 + bafilomycin. (E) Lysosomal pH measurement of HeLa cells infected with the ATCC 17978 or ATCC 17978 ΔOmpA strains stained with LysoSensor Yellow/Blue dextran for 2 h. * P =0.0164 for ATCC 17978 vs. CTL. CTL: control.

Article Snippet: Briefly, HeLa cells plated on coverslips were incubated with A. baumannii ATCC 17978 (1 x 10 7 CFU/mL) or stauroporine (10 mg/L, apopotosis inducer) for 24 h at 37 °C, 5% CO 2 .

Techniques: Infection, Labeling, Staining, Control

Journal: EMBO Molecular Medicine

Article Title: Adaptively evolved human oral actinomyces‐sourced defensins show therapeutic potential

doi: 10.15252/emmm.202114499

Figure Lengend Snippet: An inhibition‐zone‐based assay for evaluating the stability of AMSIN in H 2 O or normal mouse serum. The bacterium used was BM and the peptide dose was 0.2 nmol/well. Mean ± SD of three biological replicates is displayed. P values were obtained by Mann–Whitney U ‐test (* P < 0.05, ns: no significance; exact P values are listed in Table ). Hemolysis by AMSIN. Meucin‐18 was used as a positive control. Mean ± SD of three technical replicates is displayed. Comparison of cytotoxic effects of AMSIN and LL‐37 on HL‐60 cells. Mean ± SD of three biological replicates is displayed. P values were obtained by Student’s t ‐test or Mann–Whitney U ‐test (the latter indicated by a red asterisk; * P < 0.05, *** P < 0.001, ns: no significance; exact P values are listed in Table ). An inhibition‐zone assay showing the inability of AMSIN to bind DNA. The bacterium used was MRSA P1374 and the peptide dose was 1.0 nmol/well. Mean ± SD of three biological replicates is displayed. P values were obtained by Mann–Whitney U ‐test (ns: no significance; exact P values are listed in Table ). The effect of AMSIN on human ion channels expressed in the CNS and heart. The asterisks mark steady‐state current traces after administering 100 μM AMSIN. Peptide‐induced IL‐8 release. A549 cells were treated with AMSIN or LL‐37 for 24 h. IL‐8 levels in culture supernatants from the FBS‐containing medium ( left ) or the serum‐free medium ( right ) were measured with an ELISA, as described in Materials and Methods. Mean ± SD of three biological replicates is displayed. P values were obtained by Student’s t ‐test or Mann–Whitney U ‐test (the latter indicated by a red asterisk or ns; * P < 0.05, ** P < 0.01, *** P < 0.001, ns: no significance; exact P values are listed in Table ). Peptide‐induced MCP‐1 release. HL‐60 cells were treated with AMSIN or LL‐37 for 24 h. MCP‐1 levels in culture supernatants from the FBS‐containing medium ( left ) or the serum‐free medium ( right ) were measured with an ELISA. Mean ± SD of three biological replicates is displayed. P values were obtained by Student’s t ‐test or Mann–Whitney U ‐test (the latter indicated by a red asterisk or ns; * P < 0.05, ** P < 0.01, ns: no significance; exact P values are listed in Table ). Surface plasmon resonance (SPR)‐based assay detecting anti‐AMSIN antibodies in mouse serum using Biacore T100. Sera were taken from three AMSIN‐treated mice (Samples 1–3) and one control mouse only injected with 0.9% NaCl. The sensorgrams present data of subtracting the background SPR signals from the control. Note: the abnormal peaks at 30 and 90 s yielded by the equipment due to sample injections were artificially removed. Insect, schematic diagram of SPR experiment in which 1:100 diluted serum was used as analyte to flow over the AMSIN‐immobilized CM5 chip. Treatment of pneumococcal pneumonia. Five mice per sampling point were inoculated with SP D39 through the nasopharynx and 24 h later the animals received AMSIN or penicillin (as a positive control) at different doses (AMSIN, at the doses of 8.35, 16.7, and 33 mg per kg; penicillin at 8.35, 16.7, and 30 mg per kg) via intramuscular injection (i.m.). One day after treatment, the animals were necropsied and their lungs were removed and homogenized for the determination of the level of SP , as described in the Materials and Methods section. Each point represents the determination from a single animal and the line shows the mean log value. Mean ± SD of five to ten biological replicates is displayed. P values were obtained by Student’s t ‐test or Mann–Whitney U ‐test (the latter indicated by red asterisks; ** P < 0.01, *** P < 0.001 denotes significant reduction in CFU compared with the saline group; exact P values are listed in Table ). Survival after peritoneal infection with MRSA P1374. The survival fraction of the vehicle‐treated control group was zero out of 11 mice and the treatment group contained six mice and all survived. Source data are available online for this figure.

Article Snippet: Medium was aspirated and replaced with fresh complete DMEM supplemented with 10% FBS or serum‐free medium, both containing different concentrations of polypeptides for 24 h. Then, the supernatants were harvested for quantification of the IL‐8 amount in A549 by the Human CXCL8/IL‐8 Quantikine ELISA Kit (R&D Systems Inc., Minneapolis, MN, USA) and the MCP‐1 amount in HL‐60 by the Human MCP‐1 ELISA Kit (Boster Biological Technology Co., Ltd, Wuhan, China) as per the manufactures’ instructions.

Techniques: Inhibition, MANN-WHITNEY, Positive Control, Comparison, Enzyme-linked Immunosorbent Assay, SPR Assay, Control, Injection, Sampling, Saline, Infection

Journal: Frontiers in Oncology

Article Title: A Novel Fully-Human Potency-Matched Dual Cytokine-Antibody Fusion Protein Targets Carbonic Anhydrase IX in Renal Cell Carcinomas

doi: 10.3389/fonc.2019.01228

Figure Lengend Snippet: Biochemical characterization of IL2-XE114-TNF mut . (A) Schematic representation of the domain assembly of IL2-XE114-TNF mut in non-covalent homotrimeric format. (B) Size exclusion chromatography profile. (C) BIAcore analysis on CAIX-coated sensor chip. (D) ESI-MS profile. (E) IL2 bioactivity assay on CTLL2 cells. (F) TNF bioactivity assay on L-M fibroblasts. (G) SDS-PAGE analysis; MW, molecular weight; NR, non-reducing conditions; R, reducing conditions. (H) Flow cytometric evaluation of CAIX expression by SKRC52 cells, detected with IL2-XE114-TNF mut . (I) Differential Scanning Fluorimetry on IL2-XE114-TNF mut .

Article Snippet: CHO cells, CTLL2 cells and L-M fibroblasts were obtained from the ATCC.

Techniques: Size-exclusion Chromatography, SDS Page, Molecular Weight, Expressing

Journal: Cell

Article Title: Molecular mechanism of distinct chemokine engagement and functional divergence of the human Duffy antigen receptor

doi: 10.1016/j.cell.2024.07.005

Figure Lengend Snippet:

Article Snippet: Recombinant human CCL13 , PeproTech , Cat# 300-24.

Techniques: Virus, Recombinant, Saline, Cell Culture, Expressing, Purification, Mutagenesis, Activation Assay, Binding Assay, Phospho-proteomics, Functional Assay, Stable Transfection, Cloning, Plasmid Preparation, Software