dhrs2  (Proteintech)

Journal: Translational Pediatrics

Article Title: Untargeted lipidomics of bronchopulmonary dysplasia induced by hyperoxia exposure in rats

doi: 10.21037/tp-23-546

Figure Lengend Snippet: Alveoli in rats is impaired with hyperoxia. (A) Representative images of H&E-stained tissue slices describing alveolation 14 days after birth in rats. Arrowheads represent rat alveolar area. (B) Summary data of quantitative histomorphometric analyses: RAC. (C) Representative images of histochemical stained tissue slices describing the expression of the pulmonary SFTPD 14 days after birth in rats. The arrowheads represent cells that express SFTPD protein. (D) Quantitative analysis of immunohistochemistry results. t -test; ***, P<0.001. H&E, hematoxylin-eosin; SFTPD, surfactant-associated protein D; RAC, radial alveolar count.

Article Snippet: Use phosphate-buffered saline (PBS) containing 1.5% rabbit serum to block for 30 minutes at room temperature, then add goat anti-rabbit antibody the pulmonary surfactant-associated protein D (SFTPD; Proteintech, Wuhan, China; 1:100 dilution) to the slide and incubate overnight at 4 ℃.

Techniques: Staining, Expressing, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: DHRS2 inhibits cell growth and metastasis in ovarian cancer by downregulation of CHKα to disrupt choline metabolism

doi: 10.1038/s41419-022-05291-w

Figure Lengend Snippet: A DHRS2 protein expression levels of in OVCAR3-CON, OVCAR3-DHRS2, HO-8910-CON, HO-8910-DHRS2, SKOV3-shCON, and SKOV3-shDHRS2 cells. B Cell viability of the designated groups was detected by MTS assay. C Foci formation ability of the designated groups was determined by colony formation assay. D Cells were seeded in the upper chamber with Matrigel coating for 72 h, and cell invasion ability of the designated group was determined. Data are shown as mean values ± S.D. of independent, triplicate experiments. The asterisks (*,**,***) indicate significant differences ( p < 0.05, p < 0.01, p < 0.001, respectively).

Article Snippet: The anti-DHRS2 (PA5-25258, 1:1000) was from Thermo Fisher Scientific (Thermo Scientific, MA, USA).

Techniques: Expressing, MTS Assay, Colony Assay

Journal: Cell Death & Disease

Article Title: DHRS2 inhibits cell growth and metastasis in ovarian cancer by downregulation of CHKα to disrupt choline metabolism

doi: 10.1038/s41419-022-05291-w

Figure Lengend Snippet: A Scatter plots of all expressed genes in OVCAR3-CON and OVCAR3-DHRS2 cells. Blue color indicates upregulated genes, red indicates downregulated genes, and gray indicates non-regulated genes. The ‘regulated gene’ is defined as a gene with FDR ≤ 0.001 and abs (log2(Y/X)) ≥ 1. B Heatmap of the transcriptome changes in OVCAR3-CON and OVCAR3-DHRS2 cells ( n = 3 for each group). C Enriched KEGG pathway analysis in the transcriptome of OVCAR3-CON and OVCAR3-DHRS2 cells. D Pathway enrichment analysis in the metabolome of OVCAR3-CON and OVCAR3-DHRS2 cells. E Important features identified by partial least squares-discriminant analysis (PLS-DA). The colored boxes on the right indicate the relative concentrations of the corresponding metabolite in OVCAR3-CON and OVCAR3-DHRS2 cells.

Article Snippet: The anti-DHRS2 (PA5-25258, 1:1000) was from Thermo Fisher Scientific (Thermo Scientific, MA, USA).

Techniques:

Journal: Cell Death & Disease

Article Title: DHRS2 inhibits cell growth and metastasis in ovarian cancer by downregulation of CHKα to disrupt choline metabolism

doi: 10.1038/s41419-022-05291-w

Figure Lengend Snippet: A Oil red O staining indicating the content of LD in OVCAR3-CON, OVCAR3-DHRS2, HO-8910-CON, HO-8910-DHRS2, SKOV3-shCON, and SKOV3-shDHRS2 cells. The mRNA levels of PLA2, PLCL2, PLD1/2, CHKα, and GDPD5/6 in B OVCAR3-CON, OVCAR3-DHRS2 and C SKOV3-shCON and SKOV3-shDHRS2 cells detected by RT-qPCR. D The protein expression levels of CHKα and perilipin1 in OVCAR3-CON, OVCAR3-DHRS2, HO-8910-CON, HO-8910-DHRS2, SKOV3-shCON, and SKOV3-shDHRS2 cells. β-actin was used as a loading control. E The cellular choline content and PC/GPC ratio in OVCAR3-CON and OVCAR3-DHRS2 cells. Data are shown as mean values ± S.D. of independent, triplicate experiments. The asterisks (*,**,***) indicate significant differences ( p < 0.05, p < 0.01, p < 0.001, respectively).

Article Snippet: The anti-DHRS2 (PA5-25258, 1:1000) was from Thermo Fisher Scientific (Thermo Scientific, MA, USA).

Techniques: Staining, Quantitative RT-PCR, Expressing

Journal: Cell Death & Disease

Article Title: DHRS2 inhibits cell growth and metastasis in ovarian cancer by downregulation of CHKα to disrupt choline metabolism

doi: 10.1038/s41419-022-05291-w

Figure Lengend Snippet: A The protein expression levels of CHKα in the designated group. After treated with 10 μM choline for 3 days or transfection with CHKα, B the viability of the cells was detected by MTS, C the invasive capability of the cells was determined by cell invasion assay, and D the protein levels of p-AKT and perilipin1 were detected by Western blotting. E – G After treated with actinomycin D (5 μg/ml), CHKα mRNA levels were measured by RT-qPCR, and the percentage of remaining mRNAs in the designated group were plotted. H RIP assay showing that DHRS2 can bind to CHKα mRNA in both OVCAR3 and HO-8910 cells. Data are shown as mean values ± S.D. of independent, triplicate experiments. The asterisks (*,**,***) indicate significant differences ( p < 0.05, p < 0.01, p < 0.001, respectively).

Article Snippet: The anti-DHRS2 (PA5-25258, 1:1000) was from Thermo Fisher Scientific (Thermo Scientific, MA, USA).

Techniques: Expressing, Transfection, Invasion Assay, Western Blot, Quantitative RT-PCR

Journal: Cell Death & Disease

Article Title: DHRS2 inhibits cell growth and metastasis in ovarian cancer by downregulation of CHKα to disrupt choline metabolism

doi: 10.1038/s41419-022-05291-w

Figure Lengend Snippet: A The growth curve of OVCAR3-CON and OVCAR3-DHRS2 cells in vivo. Female BALB/c nu/nu mice were subcutaneously inoculated with OVCAR3-CON and OVCAR3-DHRS2 cells ( n = 6 per group). Tumor volume was examined every other day and shown in the graph. B At the end of the experiment, the mice were sacrificed and the tumors were separated. C Tumor mass of each group was measured and shown in the graph. D 11 C-Choline micro-PET/CT was performed and standardized uptake value (SUV) intensity was observed in the designated groups. E Representative images of Oil Red O staining in tumor tissues of the designated group. F The protein levels of DHRS2, CHKα, p-AKT, AKT, and perilipin1 in tumor tissues of the designated group. G Representative images of tumor sections in each group stained with indicated antibodies. Antibody staining is in brown and nuclear counter staining is in blue. Scatter diagram shows Histoscore for the indicated antibody staining in tumor samples. Survival analysis from TCGA dataset assessed by the Kaplan–Meier method for OC patients with high or low signature of H DHRS2 and I CHKα genes. Data are shown as mean values ± S.D. of independent, triplicate experiments. The asterisks (*,**,***) indicate significant differences ( p < 0.05, p < 0.01, p < 0.001, respectively).

Article Snippet: The anti-DHRS2 (PA5-25258, 1:1000) was from Thermo Fisher Scientific (Thermo Scientific, MA, USA).

Techniques: In Vivo, Micro-PET, Staining

Journal: Cell Death & Disease

Article Title: DHRS2 inhibits cell growth and metastasis in ovarian cancer by downregulation of CHKα to disrupt choline metabolism

doi: 10.1038/s41419-022-05291-w

Figure Lengend Snippet: A Representative images of spheroids formed by OVCAR3 cells in Matrigel (left). The invasive spheroids were featured by scattered protrusions formed on the spheroid surface, as indicated by yellow arrows. Scale bar, 50 μm. Proportion of two spheroid types in OVCAR3-CON and OVCAR3-DHRS2 cells (right) ( p < 0.05). Stable OVCAR3-CON and OVCAR3-DHRS2 cells were injected into the abdominal cavity, respectively ( n = 6 per group). B During the experiment, the body weight of the mice was determined every 3 days and shown in the graph. C Representative intraperitoneal metastatic tumors in the designated group. D , E The amount and mass of the metastatic tumors were shown in the graph. F The protein levels of DHRS2, CHKα, p-AKT, and perilipin1 in tumor tissues of the designated group. G Representative images of tumor sections in each group stained with indicated antibodies. H The representative images of DHRS2 staining in OC tissues and non-tumor tissues. The scatter diagram shows Histoscore for DHRS2 staining in the designated samples. Data are shown as mean values ± S.D. of independent, triplicate experiments. The asterisks (*,**,***) indicate significant differences ( p < 0.05, p < 0.01, p < 0.001, respectively).

Article Snippet: The anti-DHRS2 (PA5-25258, 1:1000) was from Thermo Fisher Scientific (Thermo Scientific, MA, USA).

Techniques: Injection, Staining

Journal: Cell Death & Disease

Article Title: DHRS2 inhibits cell growth and metastasis in ovarian cancer by downregulation of CHKα to disrupt choline metabolism

doi: 10.1038/s41419-022-05291-w

Figure Lengend Snippet: Left, CHKα mRNA is translated to protein, which promotes the activation of choline metabolism and AKT pathway, thereby promoting the growth and invasion of OC. Right, DHRS2 can directly bind to CHKα mRNA to accelerate its degradation. Correspondingly, CHKα protein expression is downregulated, resulting in the disruption of choline metabolism and AKT signaling. These effects lead to the inhibition of OC growth and metastasis. OC, ovarian cancer.

Article Snippet: The anti-DHRS2 (PA5-25258, 1:1000) was from Thermo Fisher Scientific (Thermo Scientific, MA, USA).

Techniques: Activation Assay, Expressing, Inhibition