Journal: iScience

Article Title: cGAS/STING sensing in dendritic cells discriminates between daptomycin sensitive and resistant Staphylococcus aureus clinical isolates

doi: 10.1016/j.isci.2026.115854

Figure Lengend Snippet: TLR2 is required for MutuDC sensing of, but not internalization of MRSA (A) Relative pHrodo labeled MRSA internalization by MutuDC over 4 h following stimulation with DapS A8819 (light blue symbols) or DapR A8817 (dark blue symbols), or media alone (white squares). Prior to stimulation, MutuDC were pre-treated for 1 h with TLR2 blocking antibody (clone T2.5; triangles with dashed lines) or media alone (circles with filled lines). Relative MRSA internalization by each DC subset is expressed as the gMFI of pHrodo. Results show the mean and (SD) of duplicates from one experiment, representative of two independent experiments. (B) Cytokine secretion (pg/mL) by MutuDC stimulated with TLR2 ligand peptidoglycan of S. aureus (PGN-SA) (10 μg/mL) or (C) DapS (A8819; light blue) or DapR (A8817; dark blue) MRSA (MOI of 10) for 18 h. MutuDC were first pre-treated with either TLR2 blocking antibody (dot-filled bars) or media alone (filled bars) as in A, or an isotype control (clone 163D3, empty bars) at 1 μg/mL. Results pooled from four (B) or three (C) independent experiments and expressed as the mean ± SEM, with each symbol (circle, square, and directional triangles) representing paired experimental replicates ( n = 3). Statistical significance determined using paired t test and reported as indicated by an ∗ when p ≤ 0.05. (D) Expression of surface activation markers by MutuDC stimulated with DapS A8819 MRSA. DC were pre-treated with TLR2 blocking antibody (black trace), isotype control (dashed red trace), and media alone (light blue shaded). Unstained control sample is shown for each marker (black dashed trace). Data shown from one experiment, representative of three independent experiments.

Article Snippet: mAB mTLR2- anti-mouse/human TLR2 , InvivoGen , Cat# mab-mtlr2; RRID: AB_763722.

Techniques: Labeling, Blocking Assay, Control, Expressing, Activation Assay, Marker

Journal: Molecular Therapy. Nucleic Acids

Article Title: Hemoglobin inhibits fibroblast-to-cardiomyocyte reprogramming via TLR2/TLR4-dependent chromatin compaction

doi: 10.1016/j.omtn.2026.102900

Figure Lengend Snippet: Hemoglobin mediates gene repression through TLR2 and TLR4 (A and B) A study was conducted to determine the effect of hemoglobin (Hb) on (A) fibroblast-to-cardiomyocyte reprogramming and (B) fibroblast gene expression. With respect to fibroblast-to-cardiomyocyte reprogramming, cardiac fibroblasts were transfected with either miR combo or a non-targeting control miR. 24 h later, the cells were incubated with vehicle, a TLR2 pharmacological inhibitor, a TLR4 pharmacological inhibitor, or a combination of both pharmacological inhibitors for 3 h. After incubation with the indicated pharmacological inhibitors, Hb was added (5 mg/mL) to the media. All media was replaced the next day. Fourteen days after miR transfection, cells were analyzed for expression of the indicated cardiomyocyte specific genes by qPCR. Expression values were normalized to the housekeeping gene Gapdh. N = 6–10. One-sample t tests were used to compare groups to the control group (∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05; ns, not significant). t test was used to determine the significance between the miR combo groups (## p < 0.01, # p < 0.05; ns, not significant). With respect to fibroblast gene expression, cardiac fibroblasts were incubated with vehicle, a TLR2 pharmacological inhibitor, a TLR4 pharmacological inhibitor, or a combination of both pharmacological inhibitors for 3 h. After incubation with the indicated pharmacological inhibitors, Hb was added (5 mg/mL) to the media. All media was replaced the next day. Expression of the indicated fibroblast-specific genes was determined by qPCR and normalized to the housekeeping gene Gapdh. N = 10. One-sample t tests were used to compare groups to the control group (∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05; ns, not significant).

Article Snippet: Hb (Millipore Sigma, H2625), TLR2 signaling inhibitor-TL2-C29 (InvivoGen, catalog no. inh-c29), and TLR4 inhibitor-CLI-095 (InvivoGen, catalog no. tlrl-cli95-4) were used.

Techniques: Gene Expression, Transfection, Control, Incubation, Expressing

Journal: Cell Reports Medicine

Article Title: Macrophage-mimetic photothermal nanotherapeutics regulate mitochondrial homeostasis and inflammatory cascades in lung ischemia-reperfusion injury

doi: 10.1016/j.xcrm.2026.102768

Figure Lengend Snippet: Preparation and characterization of Rg3@PACVs (A–C) Characterization of mPDA, Rg3@mPDA, and Rg3@PACVs using transmission electron microscopy (TEM) and dynamic light scattering (DLS) for hydrodynamic size distribution and zeta potential analysis (scale bars, 100 nm). (D) Elemental mapping of carbon (C), nitrogen (N), oxygen (O), phosphorus (P), and sulfur (S) in Rg3@PACVs (scale bars, 100 nm). (E) Fourier transform infrared spectroscopy (FTIR) spectra of mPDA, Rg3, Mac-CVs, and Rg3@PACVs. (F) The expression of macrophage-membrane markers including Toll-like receptor 2 (TLR2), integrin subunit alpha M (CD11b), C-X-C motif chemokine receptor 4 (CXCR4), receptor for advanced glycation end-products (RAGE), interleukin-6 receptor (IL-6R), and tumor necrosis factor receptor 1 (TNFR1) detected by digital western blot analysis. (G) Fluorescence of Dio-labeled Mac-CVs and Dil-labeled Rg3@mPDA after ultrasonic extrusion to form Dio/Dil-labeled Rg3@PACVs (scale bars, 1 μm). (H) Near-infrared (NIR, 808nm) images of Rg3, mPDA, Mac-CVs, and Rg3@PACVs in microcentrifuge tubes. (I and J) Temperature changes of Rg3@PACVs under different laser power intensities (I) and the photothermal effect curves of Rg3@PACVs at various concentrations (J). (K) Photothermal stability of Rg3@PACVs. (L) Size stability of Rg3@PACVs after 1-week storage at 4°C and 37°C.

Article Snippet: The target proteins included CD11b (HA722075, HUABIO, China), CXCR4 ( AWA46731 ,Abiowell, China), TLR2 (52945,Genuin Biotech, China), RAGE (ET1702-27, HUABIO), IL-6R ( AWA43078 , Abiowell), Na + /K + -ATPase (RT1412, HUABIO), PGC-1α ( AWA59146 , Abiowell), MFN2 (CSB-RA139716A0HU, CUSABIO, China), DRP1 ( HA500487 , HUABIO), phosphorylated DRP1 (Ser616) (PSH06-77, HUABIO), PI3K (Genuin Biotech), phosphorylated PI3K (U1011, Genuin Biotech), Akt (ET1609-51, HUABIO), and phosphorylated Akt (ET1607-73, HUABIO) and β-actin (EM21002, HUABIO).

Techniques: Transmission Assay, Electron Microscopy, Zeta Potential Analyzer, Fourier Transform Infrared Spectroscopy, Spectroscopy, Expressing, Membrane, Western Blot, Fluorescence, Labeling

Journal: Cell Reports Medicine

Article Title: Macrophage-mimetic photothermal nanotherapeutics regulate mitochondrial homeostasis and inflammatory cascades in lung ischemia-reperfusion injury

doi: 10.1016/j.xcrm.2026.102768

Figure Lengend Snippet:

Article Snippet: The target proteins included CD11b (HA722075, HUABIO, China), CXCR4 ( AWA46731 ,Abiowell, China), TLR2 (52945,Genuin Biotech, China), RAGE (ET1702-27, HUABIO), IL-6R ( AWA43078 , Abiowell), Na + /K + -ATPase (RT1412, HUABIO), PGC-1α ( AWA59146 , Abiowell), MFN2 (CSB-RA139716A0HU, CUSABIO, China), DRP1 ( HA500487 , HUABIO), phosphorylated DRP1 (Ser616) (PSH06-77, HUABIO), PI3K (Genuin Biotech), phosphorylated PI3K (U1011, Genuin Biotech), Akt (ET1609-51, HUABIO), and phosphorylated Akt (ET1607-73, HUABIO) and β-actin (EM21002, HUABIO).

Techniques: Recombinant, Purification, Marker, Isolation, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Apoptosis Assay, Membrane, Fluorescence, Sequencing, Gene Expression, Western Blot, Software, Real-time Polymerase Chain Reaction, Microscopy, Flow Cytometry

Journal: bioRxiv

Article Title: Liver sinusoidal endothelial cells integrate metabolic and immune signals for MAPK-dependent BMP6 regulation and hepcidin induction

doi: 10.64898/2026.05.07.723498

Figure Lengend Snippet: (A) Bmp6 mRNA expression in primary LSECs treated with 5 ng/mL LPS or 2.5 μM heme for 6 h in the presence or absence of hepatocyte-conditioned medium. (B) Bmp6 mRNA expression in primary LSECs treated with various TLR ligands: Pam3CSK4 (TLR1/2), PGN (TLR2), Poly I:C (TLR3), LPS (TLR4), FLA-ST (TLR5), FSL1 (TLR2/6), R848 (TLR7/8), and ODN (TLR9) or vehicle control (NT, non-treated) for 6 h. (C-D) Bmp6 mRNA expression in primary LSECs treated with increasing concentrations of LPS for 6 h or with 5 ng/mL LPS for 2, 4, and 6 h. (E) Bmp6 mRNA expression in LSECs treated with 2.5 μM heme or protoporphyrin IX (PPIX) for 6 h. (F-G) Bmp6 mRNA expression in primary LSECs treated with increasing concentrations of heme for 6 h or treated with 2.5 μM heme for 2, 4, or 6 h. (H) Bmp6 mRNA expression in primary LSECs pre-treated with TAK242 (5 μM) or DMSO for 1 h, followed by 5 ng/mL LPS or 2.5 μM heme treatment for 6 h. (I) Transcription factor activity analysis by RNA-seq of LSECs treated with heme with respect to untreated controls (contrast-wise), in presence of hepatocyte-conditioned medium. (J) Bmp6 mRNA expression in primary LSECs pre-treated with CHX (5 μM) or DMSO for 1 h, followed by 5 ng/mL LPS treatment for 6 h. Cell culture experiments, except those in panel A, were always conducted in the presence of hepatocyte-conditioned medium. Gene expression levels were assessed by RT-qPCR, normalized to the housekeeping gene Rpl19 , and expressed as fold change relative to vehicle-treated controls. The dashed line (ut) represents the mRNA expression of LSECs treated with the conditions shown, in the absence of LPS or heme. Data are obtained from three or four independent experiments and displayed as mean ± SD. Statistical significance: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, one-way ANOVA or Student’s t-test. CM, conditioned medium; PPIX, protoporphyrin IX; CHX, cycloheximide.

Article Snippet: The TLR ligands Pam3CSK4 (TLR2:1) (#tlrl-pms), PGN-SA (TLR2) (#tlrl-pgns2), Poly I:C (TLR3) (#tlrl-picw), FLA-ST (TLR5) (#tlrl-stfla), FSL1 (TLR2:TLR6) (tlrl-fsl), R848 (TLR7:8) (#tlrl-r848-1), ODN (TLR9) (#tlrl-1826) and the MAPK inhibitor SP600125 (#tlrl-sp60) were purchased form Invivogen and diluted in PBS (TLR ligands) or DMSO (SP600125).

Techniques: Expressing, Control, Activity Assay, RNA Sequencing, Cell Culture, Gene Expression, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Liver sinusoidal endothelial cells integrate metabolic and immune signals for MAPK-dependent BMP6 regulation and hepcidin induction

doi: 10.64898/2026.05.07.723498

Figure Lengend Snippet: (A) Bmp6 mRNA expression in primary LSECs treated with 5 ng/mL LPS or 2.5 μM heme for 6 h in the presence or absence of hepatocyte-conditioned medium. (B) Bmp6 mRNA expression in primary LSECs treated with various TLR ligands: Pam3CSK4 (TLR1/2), PGN (TLR2), Poly I:C (TLR3), LPS (TLR4), FLA-ST (TLR5), FSL1 (TLR2/6), R848 (TLR7/8), and ODN (TLR9) or vehicle control (NT, non-treated) for 6 h. (C-D) Bmp6 mRNA expression in primary LSECs treated with increasing concentrations of LPS for 6 h or with 5 ng/mL LPS for 2, 4, and 6 h. (E) Bmp6 mRNA expression in LSECs treated with 2.5 μM heme or protoporphyrin IX (PPIX) for 6 h. (F-G) Bmp6 mRNA expression in primary LSECs treated with increasing concentrations of heme for 6 h or treated with 2.5 μM heme for 2, 4, or 6 h. (H) Bmp6 mRNA expression in primary LSECs pre-treated with TAK242 (5 μM) or DMSO for 1 h, followed by 5 ng/mL LPS or 2.5 μM heme treatment for 6 h. (I) Transcription factor activity analysis by RNA-seq of LSECs treated with heme with respect to untreated controls (contrast-wise), in presence of hepatocyte-conditioned medium. (J) Bmp6 mRNA expression in primary LSECs pre-treated with CHX (5 μM) or DMSO for 1 h, followed by 5 ng/mL LPS treatment for 6 h. Cell culture experiments, except those in panel A, were always conducted in the presence of hepatocyte-conditioned medium. Gene expression levels were assessed by RT-qPCR, normalized to the housekeeping gene Rpl19 , and expressed as fold change relative to vehicle-treated controls. The dashed line (ut) represents the mRNA expression of LSECs treated with the conditions shown, in the absence of LPS or heme. Data are obtained from three or four independent experiments and displayed as mean ± SD. Statistical significance: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, one-way ANOVA or Student’s t-test. CM, conditioned medium; PPIX, protoporphyrin IX; CHX, cycloheximide.

Article Snippet: The TLR ligands Pam3CSK4 (TLR2:1) (#tlrl-pms), PGN-SA (TLR2) (#tlrl-pgns2), Poly I:C (TLR3) (#tlrl-picw), FLA-ST (TLR5) (#tlrl-stfla), FSL1 (TLR2:TLR6) (tlrl-fsl), R848 (TLR7:8) (#tlrl-r848-1), ODN (TLR9) (#tlrl-1826) and the MAPK inhibitor SP600125 (#tlrl-sp60) were purchased form Invivogen and diluted in PBS (TLR ligands) or DMSO (SP600125).

Techniques: Expressing, Control, Activity Assay, RNA Sequencing, Cell Culture, Gene Expression, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Liver sinusoidal endothelial cells integrate metabolic and immune signals for MAPK-dependent BMP6 regulation and hepcidin induction

doi: 10.64898/2026.05.07.723498

Figure Lengend Snippet: (A) Bmp6 mRNA expression in primary LSECs treated with 5 ng/mL LPS or 2.5 μM heme for 6 h in the presence or absence of hepatocyte-conditioned medium. (B) Bmp6 mRNA expression in primary LSECs treated with various TLR ligands: Pam3CSK4 (TLR1/2), PGN (TLR2), Poly I:C (TLR3), LPS (TLR4), FLA-ST (TLR5), FSL1 (TLR2/6), R848 (TLR7/8), and ODN (TLR9) or vehicle control (NT, non-treated) for 6 h. (C-D) Bmp6 mRNA expression in primary LSECs treated with increasing concentrations of LPS for 6 h or with 5 ng/mL LPS for 2, 4, and 6 h. (E) Bmp6 mRNA expression in LSECs treated with 2.5 μM heme or protoporphyrin IX (PPIX) for 6 h. (F-G) Bmp6 mRNA expression in primary LSECs treated with increasing concentrations of heme for 6 h or treated with 2.5 μM heme for 2, 4, or 6 h. (H) Bmp6 mRNA expression in primary LSECs pre-treated with TAK242 (5 μM) or DMSO for 1 h, followed by 5 ng/mL LPS or 2.5 μM heme treatment for 6 h. (I) Transcription factor activity analysis by RNA-seq of LSECs treated with heme with respect to untreated controls (contrast-wise), in presence of hepatocyte-conditioned medium. (J) Bmp6 mRNA expression in primary LSECs pre-treated with CHX (5 μM) or DMSO for 1 h, followed by 5 ng/mL LPS treatment for 6 h. Cell culture experiments, except those in panel A, were always conducted in the presence of hepatocyte-conditioned medium. Gene expression levels were assessed by RT-qPCR, normalized to the housekeeping gene Rpl19 , and expressed as fold change relative to vehicle-treated controls. The dashed line (ut) represents the mRNA expression of LSECs treated with the conditions shown, in the absence of LPS or heme. Data are obtained from three or four independent experiments and displayed as mean ± SD. Statistical significance: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, one-way ANOVA or Student’s t-test. CM, conditioned medium; PPIX, protoporphyrin IX; CHX, cycloheximide.

Article Snippet: The TLR ligands Pam3CSK4 (TLR2:1) (#tlrl-pms), PGN-SA (TLR2) (#tlrl-pgns2), Poly I:C (TLR3) (#tlrl-picw), FLA-ST (TLR5) (#tlrl-stfla), FSL1 (TLR2:TLR6) (tlrl-fsl), R848 (TLR7:8) (#tlrl-r848-1), ODN (TLR9) (#tlrl-1826) and the MAPK inhibitor SP600125 (#tlrl-sp60) were purchased form Invivogen and diluted in PBS (TLR ligands) or DMSO (SP600125).

Techniques: Expressing, Control, Activity Assay, RNA Sequencing, Cell Culture, Gene Expression, Quantitative RT-PCR