rabbit anti-activating transcription factor 6  (ABclonal Biotechnology)

Journal: Biomedicines

Article Title: Secretome of hESC-Derived MSC-like Immune and Matrix Regulatory Cells Mitigate Pulmonary Fibrosis through Antioxidant and Anti-Inflammatory Effects

doi: 10.3390/biomedicines11020463

Figure Lengend Snippet: BLM induces NOX4-mediated oxidative stress in A549 lung epithelial cells. ( a ) Representative images of immunoblotting blots revealed an increased expression of α-SMA, NOX4 and NRF2 in A549 cells exposed to indicated concentrations of BLM (1.0, 2.5, 5.0, 10.0, and 20.0 μg/mL) for 48 h. ( b ) Semi-quantitation of the relative levels of α-SMA, NOX4, and NRF2 proteins as evaluated by the relative density over GAPDH in ( a ). ( c ) Representative images of A549 cells treated with indicated concentrations of BLM for 48 h and stained with CellROX ® Orange reagent. The BLM-induced A549 injury increased ROS production, especially at the concentration of 5 μg/mL, compared to untreated cells (NC). ( d ) Mean gray value of ROS production in ( c ). ( e ) Representative images of IF exhibited more abundant HO-1, NRF2, NOX4, ATF6, α-SMA, and N-Cad proteins in A549 cells exposed to 5 μg/mL of BLM for 48 h as compared with the NC control. ( f ) Representative images of the fluorescent intensity of the relevant proteins in ( e ). Data in ( b , d , f ) represent mean ± SD from three independent experiments (N = 3), as analyzed by GraphPad prism one-way ANOVA Tukey’s multiple comparisons test. Scale bars in ( c ), 100 µm; ( e ), 10 µm.

Article Snippet: Then, the sections were stained with 1:200 rabbit monoclonal IgG Heme oxygenase-1 (HO-1) (Proteintech, 27282-1-AP), rabbit monoclonal IgG nuclear factor erythroid 2–related factor 2 (NRF2) (Proteintech, 16396-1-AP), rabbit monoclonal IgG smooth muscle actin (SMA) (Proteintech, 14395-1-AP), rabbit monoclonal IgG NADPH Oxidase 4 (NOX4) (BOSTER, BA2813), rabbit monoclonal IgG Activating Transcription Factor 6 (ATF6) (Proteintech, 24169-1-AP), rabbit monoclonal IgG N-cadherin (Proteintech, 22018-1-AP), and mouse monoclonal IgG2b TNF-α (Proteintech, 60291-1-IG) overnight at 4 °C in 5% donkey serum in PBS.

Techniques: Western Blot, Expressing, Quantitation Assay, Staining, Concentration Assay, Control

Journal: PLoS Neglected Tropical Diseases

Article Title: Genome-wide screening identified SEC61A1 as an essential factor for mycolactone-dependent apoptosis in human premonocytic THP-1 cells

doi: 10.1371/journal.pntd.0010672

Figure Lengend Snippet: Control EGFP -knockout and SEC61A1 -knockout THP-1 cells were treated with 30 ng/mL mycolactone for 6 h ( ATF4 , DDIT3 and PMAIP1 ) or 48 h ( BBC3 and BCL2L11 ). (A) The mRNA levels of ER-stress-related genes ( ATF4 and DDIT3 ) and (B) proapoptotic genes ( PMAIP1 , BBC3 and BCL2L11 ) were analyzed by real-time RT-PCR. mRNA levels are expressed relative to those of ethanol-treated EGFP- knockout or SEC61A1 -knockout THP-1 cells (n = 3). (C) The mRNA levels of unspliced, spliced and total XBP1 using control EGFP -knockout and SEC61A1 -knockout THP-1 cells treated with 30 ng/mL mycolactone for 0, 3, 6, 12, and 24 h. mRNA levels are expressed relative to those of mycolactone-treated EGFP- knockout THP-1 cells (n = 3). *: p < 0.05; **: p < 0.01; ***: p < 0.005. (D) Control EGFP -knockout and SEC61A1 -knockout THP-1 cells were treated with 30 ng/mL mycolactone for 24 h. ER stress proteins (p-eIF2α, eIF2α, p-PERK, PERK, full length ATF6, cleaved C-ATF6, and IRE1α) and SEC61A1 were evaluated by Western blotting. 1 μM thapsigargin for 24 h was included as a positive control for ER stress.

Article Snippet: The primary antibodies used were rabbit anti-caspase-3 (Cell Signaling Technology #9662, Beverly, MA, USA), rabbit anti-cleaved caspase-3 (Asp175) (Cell Signaling Technology #9661), rabbit anti-SEC61A1 (D4K2Z) (Cell Signaling Technology #14867), rabbit anti-eukaryotic initiation factor (eIF2)α (D7D3) (Cell Signaling Technology #5325), rabbit anti-phospho-eIF2α (Ser51) (D9D8) (Cell Signaling Technology #3398), rabbit anti- protein kinase-like ER kinase (PERK) (C33E10) (Cell Signaling Technology #3192), rabbit anti-phospho-PERK (Thr980) (16F8) (Cell Signaling Technology #3179), rabbit anti-activating transcription factor (ATF6) (D4Z8V) (Cell Signaling Technology #65880), rabbit anti-inositol requiring enzyme-1 (IRE1)α (14C10) (Cell Signaling Technology #3294) and mouse anti-β-actin (Sigma-Aldrich #5441).

Techniques: Control, Knock-Out, Quantitative RT-PCR, Western Blot, Positive Control