Additional file 1 of Ventilator-induced lung-injury in mouse models: Is there a trap?

Additional file 1. Supplemental table 1

Additional file 1 of Ventilator-induced lung injury results in oxidative stress response and mitochondrial swelling in a mouse model

Additional file 1. Figure S1: Whole lung sections. Shown are whole lung sections from SB and different tidal volume ventilated mice. The boxes drawn show areas represented in Fig. 3. Width of box is 450 µm and height is 330 µ

Additional file 2 of Ventilator-induced lung injury results in oxidative stress response and mitochondrial swelling in a mouse model

Additional file 2. Figure S2: Junctional gene sets negatively enriched due to mechanical ventilation. APICAL_JUNCTION gene set is negatively correlated due to mechanical ventilation (A). CLDN4 expression is increased due to mechanical ventilation. Standard deviation of the means is shown. Significant difference from the SB controls are shown (P ≤ 0.01 = **; P ≤ 0.001 = ***

Sensitive detection of lysosomal membrane permeabilization by lysosomal galectin puncta assay

Lysosomal membrane permeabilization (LMP) contributes to tissue involution, degenerative diseases, and cancer therapy. Its investigation has, however, been hindered by the lack of sensitive methods. Here, we characterize and validate the detection of galectin puncta at leaky lysosomes as a highly sensitive and easily manageable assay for LMP. LGALS1/galectin-1 and LGALS3/galectin-3 are best suited for this purpose due to their widespread expression, rapid translocation to leaky lysosomes and availability of high-affinity antibodies. Galectin staining marks individual leaky lysosomes early during lysosomal cell death and is useful when defining whether LMP is a primary or secondary cause of cell death. This sensitive method also reveals that cells can survive limited LMP and confirms a rapid formation of autophagic structures at the site of galectin puncta. Importantly, galectin staining detects individual leaky lysosomes also in paraffin-embedded tissues allowing us to demonstrate LMP in tumor xenografts in mice treated with cationic amphiphilic drugs and to identify a subpopulation of lysosomes that initiates LMP in involuting mouse mammary gland. The use of ectopic fluorescent galectins renders the galectin puncta assay suitable for automated screening and visualization of LMP in live cells and animals. Thus, the lysosomal galectin puncta assay opens up new possibilities to study LMP in cell death and its role in other cellular processes such as autophagy, senescence, aging, and inflammation.</p

Additional file 1: of Azithromycin induces epidermal differentiation and multivesicular bodies in airway epithelia

Figure S1. Azm treatment of BCi-NS1.1 cells leads to increased vesicle formation Transmission electron microscope images show that BCi-NS1.1 cells treated with Azm have substantially more vesicle formation than the untreated controls. Left scale bars are 10.0 μm and right scale bars are 1.0 μm. (TIF 18391 kb

Additional file 2: of Azithromycin induces epidermal differentiation and multivesicular bodies in airway epithelia

Figure S2. Azm induced MVB and LB formation. A) BCi-NS1.1 cells differentiated in ALI cultures treated with Azm showed a marked increase in MVB and LB formations. Shown are two different cross sectional TEM images of transwell filters. Scale bars are from left 10.0, 2.0, 1.0 and 1.0 μm. B) Treating differentiated VA10 cells with a clinical formulation of Azm (Zithromax) also resulted in increased MVB and LB formations. Top scale bars are 5.0 μm and bottom scale bars are 1.0 μm. C) Differentiated BCi-NS1.1 cells showed similar MVB and LB formations after treatment with Azm (Zithromax). Top scale bars are 5.0 μm and bottom scale bars are 1.0 μm. (TIF 33306 kb