Journal: Nanophotonics

Article Title: Repeated porphyrin lipoprotein-based photodynamic therapy controls distant disease in mouse mesothelioma via the abscopal effect

doi: 10.1515/nanoph-2021-0241

Figure Lengend Snippet: Figure 3: PDT reduced calreticulin expression in irradiated tumors. Dual AE17-OVA+ tumor bearing mice were treated once with either PBS, PLP, PBS + laser irradiation, or PLP + laser irradiation. Twenty-four hours after irradiation, irradiated and nonirradiated tumors were collected and processed for immunohistochemistry. (A) Whole slide scan of calreticulin stained tumors in mice for irradiated tumors on the left flank and nonirradiated tumors on the right flank. Scale bars represent 5 mm. (B) Quantification of the whole tissue slide by Halo (n = 3–4 per group) for the left (LT) and right (RT) tumors. Data are mean ± standard deviation. Statistical significance was determined using a one-way ANOVA followed by a post-hoc Tukey test. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: An anti-calreticulin antibody was purchased from Novus Biologicals (Calreticulin Antibody, NB600-103).

Techniques: Expressing, Irradiation, Immunohistochemistry, Staining, Standard Deviation

Journal: Immunology

Article Title: Acute infection of mice with Clostridium difficile leads to eIF2α phosphorylation and pro-survival signalling as part of the mucosal inflammatory response.

doi: 10.1111/imm.12122

Figure Lengend Snippet: Figure 6. Assessment of the unfolded protein response in Clostridium difficile-infected (CDI) mice. Protein lysates from the caeca and colons of untreated and C. difficile-infected mice were used to evaluate the phosphorylation of eIF2a (n = 6 pairs) (a), and the expression levels of BiP (n = 6 pairs) (d), P58IPK (n = 6 pairs) (e) and calreticulin (n = 6 pairs) (f). In each case, the panel on the left shows the image of the immunoblot for the evaluated molecule for three pairs of caeca and colons and the bar graph on the right depicts the mean SEM of the response for all the six evaluated pairs. The expression levels of Gadd34 and Wars were determined by quantitative RT-PCR (n = 12 pairs). Each bar represents the mean SEM of the relative expression of the depicted gene (b). Conventional RT-PCR was used to determine the lack of Xbp1 splicing (n = 12 pairs, of which 6 are shown) (c). A P-value ≤0.05 indicates a significant difference between the untreated and CDI samples and is marked with a ⋆.

Article Snippet: Immunoblotting antibodies against b-actin (clone 13E5), calreticulin, phospho-eIF2a (clone 119A11), eIF2a (clone L57A5), GAPDH (clone 14C10), P58IPK (clone C56E7), phospho-AKT (clone D9E), AKT (clone C67E7), phospho-STAT3 (clone D3A7) and STAT3 (clone 79D7) were obtained from Cell Signaling Technology (Danvers, MA).

Techniques: Infection, Phospho-proteomics, Expressing, Western Blot, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction

Journal: BMC ophthalmology

Article Title: Proteomic analysis of retinal pigment epithelium cells after exposure to UVA radiation.

doi: 10.1186/s12886-019-1151-9

Figure Lengend Snippet: Fig. 3 Identification of the differentially expressed proteins between the control and UVA irradiated ARPE-19 cells by 2-DE. a ARPE-19 cells irradiated with 10 J/cm2 UVA, followed by harvesting the cells and cell lysates. A total of 300 μg protein was then subjected to 2-DE and visualized by CBR staining. PDQuest image analysis software (Bio-Rad) was employed to explore differential expression through differential protein spotting. b Validation of identified selected proteins from 2-DE by western blot analysis. Some identified proteins include GRP78, CALR, PDI, NPM, Hsp27, ATP synthase, GAPDH, and PRDX1. β-actin was used as the loading control

Article Snippet: We purchased antibodies against Heat shock protein 27 (Hsp27), nucleophosmin (NPM), ATP synthase, and GAPDH from Epitomics (Burlingame, CA, USA) and antibodies against glucose-related protein 78 (GRP78), calreticulin (CALR), protein disulfide isomerase (PDI), cleaved-ATF6, ATF4, and PRDX1 antibodies were purchased from ProteinTech Group (Chicago, IL, USA).

Techniques: Control, Irradiation, Staining, Software, Quantitative Proteomics, Biomarker Discovery, Western Blot

Journal: eLife

Article Title: Biallelic TANGO1 mutations cause a novel syndromal disease due to hampered cellular collagen secretion

doi: 10.7554/eLife.51319

Figure Lengend Snippet: Immunofluorescence images of U2OS cells, transiently transfected with WT TANGO1-HA or Ex8-HA. Representative images of three independent experiments. ( A ) Cells were probed with anti-HA (green), anti-Sec16A (red) and anti-Calreticulin (blue) antibodies. Scale bar 10 μm. ( B ) Merged images of TANGO1-HA or Ex8-HA (green) and sec16A (red) and a plot comparing Manders’ overlap coefficient of HA with sec16A in TANGO1- or Ex8-expressing U2OS cells. ( C ) Merged images of TANGO1-HA or Ex8-HA (green) with calreticulin (red) and a plot comparing Manders’ overlap coefficient of HA and calreticulin in TANGO1- or Ex8-expressing U2OS cells. Student’s t test was performed to compare the Manders’ overlap coefficients, p values are shown.

Article Snippet: Antibodies used in Western blotting and immunofluorescence microscopy were as follows: TANGO1 (Sigma-Aldrich); beta-tubulin (Sigma-Aldrich); calreticulin (Novus Biologicals, Centennial, Colorado, USA); Calnexin (Abcam, Cambridge, United Kingdom); antitrypsin Ab-1 (NeoMarkers, Fremont, California, USA); collagen I and collagen IV (Abcam, Cambridge, United Kingdom); collagen XII (Santa Cruz Biotechnology, Dallas, Texas, USA); Sec16A (Sigma-Aldrich); rat hemagglutinin (Roche) or mouse hemagglutinin (Santa Cruz Biotechnology, Dallas, Texas, USA).

Techniques: Immunofluorescence, Transfection, Expressing