Journal: bioRxiv

Article Title: Phosphoarginine modulates oligomerization and repressor activity of mycobacterial ClpC2

doi: 10.64898/2026.06.30.735635

Figure Lengend Snippet: A) Diagram of the DNA template used for in vitro transcription studies, including a σ70-dependent E. coli promoter, clpC2 operator, and sequence encoding a F30-broccoli RNA aptamer. B) Transcription of the broccoli aptamer was monitored by the increase in fluorescence over time (green trace). Inclusion of 6 µM Mtb ClpC2 decreased transcription (red trace). DFHBI-1T alone produced minimal fluorescence (gray trace). C) Transcription from a template incorporating a scrambled operator sequence was similar in the absence (cyan) or presence (blue) of 6 µM Mtb ClpC2. By contrast, 2 µM rifampin completely inhibited transcription (orange trace). D) Transcriptional rate was measured as a function of Msm ClpC2 concentration, using templates with scrambled or intact operator sequences. E) Transcription was measured in the presence of the indicated concentration of Mtb ClpC2, or with the additional inclusion of 500 µM pArg, 10 µM pArg casein, 10 µM unphosphorylated casein, 1.2-fold molar excess Ruf or 1.2-fold molar excess CymA. F). Transcriptional repression is shown for the indicated concentration of wild-type Mtb ClpC2, and for Mtb ClpC2 variants incorporating R56A or T256A mutations. Error bars reflect standard deviation of ≥3 replicates. Statistical significance was determined by ordinary one-way ANOVA followed by Dunnett’s multiple comparison test to compare each condition to ClpC2 alone (blue bar) at a given concentration. Multiplicity-adjusted p -values are indicated as follows: * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001. Non-significant comparisons are not shown.

Article Snippet: In a separate tube, 200 nM of template DNA was incubated in buffer with or without ClpC2, pArg casein (10 μM), casein (10 μM), pArg (500 μM), Ruf (1.2-fold molar excess), or CymA (1.2-fold molar excess) at 37°C for 15 min. DNA mixtures and holoenzyme were mixed and incubated at 37°C for 15 min, followed by addition of 100 nM of DFHBI-1T (MedChemExpress).

Techniques: In Vitro, Sequencing, Fluorescence, Produced, Concentration Assay, Standard Deviation, Comparison

Journal: Bioactive Materials

Article Title: Inhalable PD-L1-engineered hybrid cellular vesicles suppress excessive neutrophil activation and restore mitochondrial homeostasis to alleviate ischemia–reperfusion lung injury and pneumonia

doi: 10.1016/j.bioactmat.2026.03.024

Figure Lengend Snippet: Res-PD-L1@nmEVs Effectively Attenuates MRSA-Induced Pneumonia (A-B) Rats with MRSA-induced pneumonia received three bronchial nebulization treatments over one week with different formulations (Res, nEVs, PD-L1@mEVs, PD-L1@nmEVs, or Res-PD-L1@nmEVs). (A) Representative H&E-stained lung sections and (B) corresponding lung injury scores are shown (n = 5). (C) TUNEL staining of lung tissues to assess apoptosis. (D) Representative micro-CT images of anesthetized rats. (E-G) Flow cytometric analysis of immune cell proportions in lung single-cell suspensions: CD8 + T cells (E), neutrophils (F), and classical monocytes (G). (H-J) Plasma levels of inflammatory cytokines IL-6 (H), IL-1β (I), and TNF-α (J) (n = 5). (K) Immunofluorescence staining of tight junction proteins Occludin (green) and ZO-1 (red) in lung tissues (nuclei stained with DAPI). Scale bar: 50 μm. (L-N) Pulmonary function parameters: lung compliance (L), airway resistance (M), and oxygenation index (N) (n = 4). ∗ vs. Sham; # vs. MRSA; & vs. MRSA + PD-L1@nmEVs, p < 0.05.

Article Snippet: Apoptosis detection was performed using the TUNEL reaction mixture (G1502, Servicebio) after fixation and permeabilization, with incubation at 37 °C for 1 h in the dark.

Techniques: Staining, TUNEL Assay, Micro-CT, Single Cell, Clinical Proteomics, Immunofluorescence