Journal: Microorganisms

Article Title: Precipitation of Magnetic Iron Oxide Induced by Sporosarcina pasteurii Cells

doi: 10.3390/microorganisms9020331

Figure Lengend Snippet: Magnetic microbial-induced precipitation (MIP) induced by whole cells of Sporosarcina pasteurii . ( A ) Growth curve of S. pasteurii. ( B ) Schematic diagram of magnetic MIP induced by S. pasteurii . ( C ) Curve of residual FeCl 2 in the supernatant during magnetic MIP. All values were determined in three independent biological replicates.

Article Snippet: Sporosarcina pasteurii (ATCC 11859) was used in the MIP experiment in this study.

Techniques:

Journal: Pathology and Oncology Research

Article Title: A Novel BRD Family PROTAC Inhibitor dBET1 Exerts Great Anti-Cancer Effects by Targeting c-MYC in Acute Myeloid Leukemia Cells

doi: 10.3389/pore.2022.1610447

Figure Lengend Snippet: BRD4 is a promising therapeutic target for AML. (A) The expression of BRD4 was characterized in different cancer cell lines. High expression of BRD4 was observed in AML cell lines. (B) In a cohort of adult patient with AML ( n = 148), patients with high expression of BRD4 was significantly associated with poorer overall survival (OS) ( p = 0.044). (C) Expression of BRD4 was evaluated in four AML cell lines: NB4 (PML-RARa), Kasumi (AML1-ETO), THP-1 (MLL-AF9), and MV4-11 (MLL-AF4), cells, BRD4 was positively expressed in all cell lines observed using Western blot. (D) GAPDH served as an internal control. Densities of protein bands of BRD4 in each cell line were quantified by ImageJ and normalized to the densities of GAPDH bands. (E) Expressions of BRD2 and BRD3 were evaluated in the same four AML cell lines: NB4 (PML-RARa), Kasumi (AML1-ETO), THP-1 (MLL-AF9), and MV4-11 (MLL-AF4), cells, BRD2 and BRD3 were also positively expressed in all cell lines observed using Western blot. (F) The cytotoxic effects of dBET1 were determined using CCK-8 assay. The IC 50 was estimated for each cell line.

Article Snippet: Primary antibodies against the following proteins were used: BRD2(cat:5848s, 1:1,000, Cell Signaling Technology), BRD3(cat: 11859-1-AP, 1:1,000, Proteintech), BRD4 (cat: 13440s, 1:1,000, Cell Signaling Technology), CRBN (cat: HPA045910, Sigma-Aldrich), c-MYC (cat: 9402, 1:1,000, Cell Signaling Technology), PARP (Cat: 9542, 1:1,000, Cell Signaling Technology).

Techniques: Expressing, Western Blot, CCK-8 Assay

Journal: Pathology and Oncology Research

Article Title: A Novel BRD Family PROTAC Inhibitor dBET1 Exerts Great Anti-Cancer Effects by Targeting c-MYC in Acute Myeloid Leukemia Cells

doi: 10.3389/pore.2022.1610447

Figure Lengend Snippet: BRD2, BRD3, and BRD4 are specific targets of dBET1 in a CRBN-dependent manner. (A) After treatment with dBET1, the expression of BRD2, BRD3, and BRD4 was determined using Western blotting in NB4, Kasumi, MV4-11, and THP-1 cells. Compared with that in the control, the expression of BRD2, BRD3, and BRD4 was decreased in a dose-dependent manner. (B) Since dBET1 is a CRBN-based BET degrader, we knocked down CRBN using shRNA and found that downregulation of CRBN significantly increased the resistance of NB4 and MV4-11 cells to dBET1 (top panel). In contrast, overexpression of CRBN increased the sensitivity ofNB4 and MV4-11 cells to dBET1 (bottom panel). (C) Since dBET1-mediated degradation of BRD2, BRD3, and BRD4 was through a ubiquitin–proteasome system, we used the proteasome inhibitor, (R)-MG132, which inhibited the degradation of BRD2, BRD3, and BRD4 in a dose-dependent manner. KD: knock down, OE: overexpression, scr: scramble.

Article Snippet: Primary antibodies against the following proteins were used: BRD2(cat:5848s, 1:1,000, Cell Signaling Technology), BRD3(cat: 11859-1-AP, 1:1,000, Proteintech), BRD4 (cat: 13440s, 1:1,000, Cell Signaling Technology), CRBN (cat: HPA045910, Sigma-Aldrich), c-MYC (cat: 9402, 1:1,000, Cell Signaling Technology), PARP (Cat: 9542, 1:1,000, Cell Signaling Technology).

Techniques: Expressing, Western Blot, shRNA, Over Expression