Journal: Cancer research

Article Title: Hedgehog signaling regulates metabolism and polarization of mammary tumor-associated macrophages

doi: 10.1158/0008-5472.CAN-20-1723

Figure Lengend Snippet: A. Schematic of O-GlcNAcylation. Ogt adds O-GlcNAc to target proteins from the substrate UDP-GlcNAc, and Oga removes O-GlcNAc moieties. B. Immunoblot for all cellular O-GlcNAcylated proteins (using RL2 antibody) in M2 macrophages (BMDMs) +/− Hhi. C. Protein (Immunoblot) and transcript (qPCR) levels for Ogt in M2 macrophages (BMDMs) +/− Hhi. D. ChIP was performed in macrophages (skewed from RAW264.7 cells) for Gli1 binding to the Ogt promoter followed by qPCR with primers specific for Ogt promoter. Graph shows qPCR results with signal relative to input with beads subtracted. E. Immunoblots for cellular O-GlcNAcylated proteins (using RL2 antibody) in M2 macrophages sorted from 4T1 tumors and EMT6 tumors of mice treated with DMSO or Smo-i. F. Immunoblots for Ogt in tumor infiltrating M2 macrophages sorted from 4T1 and EMT6 tumors of mice treated with DMSO or Smo-i *P<0.05. These lysates were also used for data depicted in Figure 8C . As such the same image for β-actin loading control is used.

Article Snippet: RAW264.7 and 4T1 cells were obtained from ATCC, while the EMT6 cells were a gift from Dr. Sophia Ran, Southern Illinois University.

Techniques: Western Blot, Binding Assay, Control

Journal: OncoTargets and therapy

Article Title: Decreased expression of SLC 39A14 is associated with tumor aggressiveness and biochemical recurrence of human prostate cancer

doi: 10.2147/OTT.S103640

Figure Lengend Snippet: Decreased expression of SLC39A14 protein and mRNA in human PCa tissues. Notes: ( A ) SLC39A14 protein was mainly localized in the membrane and cytoplasm of prostate cells in adjacent noncancerous prostate tissues. Red arrows show strong positive immunostrainings. Magnification, ×400. ( B ) SLC39A14 protein was weakly expressed in cancer cells in PCa tissues. Magnification, ×400. ( C ) Statistical analysis revealed that SLC39A14 protein expression levels in human PCa tissues were significantly lower than those in adjacent noncancerous prostate tissues ( P <0.01). ( D ) SLC39A14 mRNA expression levels in human PCa tissues were also lower than those in adjacent noncancerous prostate tissues based on the Taylor dataset ( P <0.01). ( E ) SLC39A14 mRNA expression levels in human PCa tissues were also lower than those in adjacent noncancerous prostate tissues based on our clinical samples ( P <0.01). Abbreviations: PCa, prostate cancer; SLC39A14, solute carrier family 39, member 14.

Article Snippet: Rabbit polyclonal ZIP14 antibody (ProSci incorporated, Poway, CA, USA) was used in this study.

Techniques: Expressing, Membrane

Journal: OncoTargets and therapy

Article Title: Decreased expression of SLC 39A14 is associated with tumor aggressiveness and biochemical recurrence of human prostate cancer

doi: 10.2147/OTT.S103640

Figure Lengend Snippet: Kaplan–Meier curves of patients with PCa based on SLC39A14 mRNA expression. Notes: ( A ) There was a significant difference in BCR-free survival between patients with high and low SLC39A14 mRNA expression ( P =0.017). ( B ) There was no significant difference in overall survival between patients with high and low SLC39A14 mRNA expression ( P =0.148). Abbreviations: BCR, biochemical recurrence; PCa, prostate cancer; SLC39A14, solute carrier family 39, member 14.

Article Snippet: Rabbit polyclonal ZIP14 antibody (ProSci incorporated, Poway, CA, USA) was used in this study.

Techniques: Expressing

Journal: OncoTargets and therapy

Article Title: Decreased expression of SLC 39A14 is associated with tumor aggressiveness and biochemical recurrence of human prostate cancer

doi: 10.2147/OTT.S103640

Figure Lengend Snippet: Downregulation of SLC39A14 promotes cell proliferation of LNCaP cells in vitro. Notes: ( A ) Western blot analysis showed that SLC39A14 protein expression was significantly upregulated by the transfection of SLC39A14 plasmid (en-SLC39A14 or en-con), but was significantly downregulated by the transfection of SLC39A14 siRNA (si-SLC39A14 or si-con). ( B ) CCK-8 assay indicated that the cell viability of LNCaP cells with overexpression of SLC39A14 was significantly lower than those of control vector-transfected cells. ( C ) CCK-8 assay indicated that the cell viability of LNCaP cells with knockdown of SLC39A14 dramatically promoted the cell viability. Abbreviations: CCK-8, Cell Counting Kit-8; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; SLC39A14, solute carrier family 39, member 14; OD, optical density.

Article Snippet: Rabbit polyclonal ZIP14 antibody (ProSci incorporated, Poway, CA, USA) was used in this study.

Techniques: In Vitro, Western Blot, Expressing, Transfection, Plasmid Preparation, CCK-8 Assay, Over Expression, Control, Knockdown, Cell Counting

Journal: OncoTargets and therapy

Article Title: Decreased expression of SLC 39A14 is associated with tumor aggressiveness and biochemical recurrence of human prostate cancer

doi: 10.2147/OTT.S103640

Figure Lengend Snippet: Downregulation of SLC39A14 promotes invasion of LNCaP cells in vitro. Notes: Transwell assays clearly revealed that downregulation of SLC39A14 significantly enhanced the invasion activity of LNCaP cells compared to that of control cells at 24 hours after the transfection, while overexpression of SLC39A14 dramatically reduced the cell invasion. Cell invasion of LNCaP cells transfected with en-con ( A ), en-SLC39A14 ( B ), si-con ( C ) and si-SLC39A14 ( D ). ( E ) The number of invasive cells after the transfection of en-con and en-SLC39A14. ( F ) The number of invasive cells after the transfection of si-con and si-SLC39A14. Abbreviation: SLC39A14, solute carrier family 39, member 14.

Article Snippet: Rabbit polyclonal ZIP14 antibody (ProSci incorporated, Poway, CA, USA) was used in this study.

Techniques: In Vitro, Activity Assay, Control, Transfection, Over Expression

Journal: OncoTargets and therapy

Article Title: Decreased expression of SLC 39A14 is associated with tumor aggressiveness and biochemical recurrence of human prostate cancer

doi: 10.2147/OTT.S103640

Figure Lengend Snippet: Downregulation of SLC39A14 promotes migration of LNCaP cells in vitro. Notes: ( A ) Cell migration of LNCaP cells transfected with en-con, en-SLC39A14, si-con and si-SLC39A14. ( B ) Inhibition rate of LNCaP cells after the transfection of en-con and en-SLC39A14. ( C ) Inhibition rate of LNCaP cells after the transfection of si-con and si-SLC39A14. Wound-healing assays demonstrated that downregulation of SLC39A14 significantly enhanced the migration activity of LNCaP cells compared to that of control cells at 24 hours after the transfection, while overexpression of SLC39A14 dramatically reduced the cell migration. Abbreviation: SLC39A14, solute carrier family 39, member 14.

Article Snippet: Rabbit polyclonal ZIP14 antibody (ProSci incorporated, Poway, CA, USA) was used in this study.

Techniques: Migration, In Vitro, Transfection, Inhibition, Activity Assay, Control, Over Expression

Journal: Cureus

Article Title: Lack of Effect of Omecamtiv Mecarbil on Smooth Muscle Myosin Phosphatase Target Subunit-1 (MYPT1) Phosphorylation and Hemodynamics in Rats

doi: 10.7759/cureus.87854

Figure Lengend Snippet: Myosin phosphatase target subunit 1 (MYPT1) phosphorylation at Thr696 or Thr695 was assessed in protein fractions extracted from the aortic rings incubated with phenylephrine (10 -5 mol/L) alone or in combination with omecamtiv mecarbil (10 -5 mol/L). The upper two panels show immunoblot signals detected using antibodies targeting p-MYPT (Thr696), p-MYPT (Thr695), t-MYPT, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Note that phenylephrine (10 -5 mol/L) alone and in combination with omecamtiv mecarbil (10 -5 mol/L) induced similar levels of MYPT1 phosphorylation at both sites.

Article Snippet: Some rings were incubated for 15 minutes with omecamtiv mecarbil (CK-1827452, MedChemExpress, Monmouth Junction, USA, 10 -5 and 10 -6 mol/L, n = 7 each), whereas control rings received no additional compound (n = 9).

Techniques: Phospho-proteomics, Incubation, Western Blot

Journal: Cureus

Article Title: Lack of Effect of Omecamtiv Mecarbil on Smooth Muscle Myosin Phosphatase Target Subunit-1 (MYPT1) Phosphorylation and Hemodynamics in Rats

doi: 10.7759/cureus.87854

Figure Lengend Snippet: The time course of heart rate (bpm) and mean arterial pressure (mmHg) is shown following intravenous administration of omecamtiv mecarbil (10 -5 mol/L) or equivalent volume of vehicle (time control) until a 30-minute period. Note that neither omecamtiv mecarbil nor the vehicle altered mean arterial pressure or heart rate in rats under general anesthesia induced with a mixture of α-chloralose and urethane.

Article Snippet: Some rings were incubated for 15 minutes with omecamtiv mecarbil (CK-1827452, MedChemExpress, Monmouth Junction, USA, 10 -5 and 10 -6 mol/L, n = 7 each), whereas control rings received no additional compound (n = 9).

Techniques: Control