antibody for plpp1  (Novus Biologicals)

Journal: International Journal of Molecular Sciences

Article Title: Z-Ligustilide Combined with Cisplatin Reduces PLPP1-Mediated Phospholipid Synthesis to Impair Cisplatin Resistance in Lung Cancer

doi: 10.3390/ijms242317046

Figure Lengend Snippet: Z-ligustilide+cisplatin reduced the phospholipid contents and upregulated PLPP1 expression. ( A ) Heat map of the differential lipids in A549, A549/DDP cells and A549/DDP cells for Z-ligustilide+cisplatin. ( B ) Pathway enrichment analysis. ( C ) Relative enrichment of each lipid class. *** p < 0.001. ( D ) GSEA analysis of the RNA-seq data. The glycerophospholipid metabolism pathway–targeted genes show significant overlap. ( E ) Heat map for RNA–Seq analysis of significantly differentially expressed genes in A549 cells vs. A549/DDP cells vs. A549/DDP cells treated with Z-ligustilide+cisplatin. Red box represented the target gene. ( F ) Protein level of PLPP1 determined via Western blot.

Article Snippet: The primary antibody for PLPP1 (known as PPAP2A, Cat. No. NBP1-59011; Novus Biologicals, Colorado, USA) was used for immunohistochemical staining, and the dilution ratio of the PLPP1 antibody was 1:200.

Techniques: Expressing, RNA Sequencing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Z-Ligustilide Combined with Cisplatin Reduces PLPP1-Mediated Phospholipid Synthesis to Impair Cisplatin Resistance in Lung Cancer

doi: 10.3390/ijms242317046

Figure Lengend Snippet: PLPP1 expression was low and associated with poor prognosis in lung cancer. ( A ) PLPP1 expression was detected in LUAD, LUSC, and corresponding normal tissues. LUAD (num (T) = 483; num (N) = 347). LUSC (num (T) = 486; num (N) = 338). * p < 0.05. ( B ) Representative images of the immunohistochemical staining of PLPP1 in lung cancer samples. Scale bars, 50 μm. −: negative expression; +: lower expression; ++: moderate expression; +++: high expression. The score criteria are described in detail in Materials and Methods. ( C ) The association between the PLPP1 protein expression and overall survival in patients with lung cancer. Low PLPP1 expression (n = 41), high PLPP1 expression (n = 35). * p < 0.05. ( D ) The correlation between PLPP1 expression and overall survival in 1451 lung cancer patients. ( E ) The correlation between PLPP1 expression and overall survival in 116 lung cancer patients administered chemotherapy. ( F ) The correlation between PLPP1 expression and overall survival in 268 lung cancer patients without chemotherapy.

Article Snippet: The primary antibody for PLPP1 (known as PPAP2A, Cat. No. NBP1-59011; Novus Biologicals, Colorado, USA) was used for immunohistochemical staining, and the dilution ratio of the PLPP1 antibody was 1:200.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: International Journal of Molecular Sciences

Article Title: Z-Ligustilide Combined with Cisplatin Reduces PLPP1-Mediated Phospholipid Synthesis to Impair Cisplatin Resistance in Lung Cancer

doi: 10.3390/ijms242317046

Figure Lengend Snippet: Z-ligustilide+cisplatin induced the reversal of resistance mediated by PLPP1. ( A ) Protein expression of PLPP1 in shCtrl and PLPP1 knockdown A549/DDP and H460/DDP cells, respectively. ( B ) Cell viability was measured in shCtrl and PLPP1 knockdown A549/DDP and H460/DDP cells with or without the Z-ligustilide and cisplatin combination treatment, *** p < 0.001. ( C ) Relative protein expressions in shCtrl and PLPP1 knockdown A549/DDP and H460/DDP cells with or without the Z-ligustilide and cisplatin combination treatment.

Article Snippet: The primary antibody for PLPP1 (known as PPAP2A, Cat. No. NBP1-59011; Novus Biologicals, Colorado, USA) was used for immunohistochemical staining, and the dilution ratio of the PLPP1 antibody was 1:200.

Techniques: Expressing, Knockdown

Journal: International Journal of Molecular Sciences

Article Title: Z-Ligustilide Combined with Cisplatin Reduces PLPP1-Mediated Phospholipid Synthesis to Impair Cisplatin Resistance in Lung Cancer

doi: 10.3390/ijms242317046

Figure Lengend Snippet: Z-ligustilide+cisplatin induced the inactivation of AKT by inhibiting PLPP1-mediated phospholipid synthesis. ( A ) The relative enrichments of each lipid class in shCtrl and PLPP1 knockdown A549/DDP cells with or without the Z-ligustilide and cisplatin combination treatment. ns, not significant, *** p < 0.001. ( B ) Protein expression of p-AKT in shCtrl and PLPP1 knockdown A549/DDP and H460/DDP cells with or without the Z-ligustilide and cisplatin combination treatment. ( C , D ) PI3K kinase activity assay and PIP 3 level measurements in shCtrl and PLPP1 knockdown A549/DDP and H460/DDP cells with or without the Z-ligustilide and cisplatin combination treatment. ns, not significant, ** p < 0.01 and *** p < 0.001.

Article Snippet: The primary antibody for PLPP1 (known as PPAP2A, Cat. No. NBP1-59011; Novus Biologicals, Colorado, USA) was used for immunohistochemical staining, and the dilution ratio of the PLPP1 antibody was 1:200.

Techniques: Knockdown, Expressing, Kinase Assay

Journal: International Journal of Molecular Sciences

Article Title: Z-Ligustilide Combined with Cisplatin Reduces PLPP1-Mediated Phospholipid Synthesis to Impair Cisplatin Resistance in Lung Cancer

doi: 10.3390/ijms242317046

Figure Lengend Snippet: Z-ligustilide+cisplatin reduced the phospholipid contents and upregulated PLPP1 expression. ( A ) Heat map of the differential lipids in A549, A549/DDP cells and A549/DDP cells for Z-ligustilide+cisplatin. ( B ) Pathway enrichment analysis. ( C ) Relative enrichment of each lipid class. *** p < 0.001. ( D ) GSEA analysis of the RNA-seq data. The glycerophospholipid metabolism pathway–targeted genes show significant overlap. ( E ) Heat map for RNA–Seq analysis of significantly differentially expressed genes in A549 cells vs. A549/DDP cells vs. A549/DDP cells treated with Z-ligustilide+cisplatin. Red box represented the target gene. ( F ) Protein level of PLPP1 determined via Western blot.

Article Snippet: The following primary antibodies for BAX (Cat. No. 50599-2-Ig; Proteintech Technology, Wuhan, China), PLPP1 (PPAP2A, which was used in western blot experiments) (Cat. No. 17967-1-AP; Proteintech Technology, Wuhan, China) Cyclin A2 (Cat. No. 18202-1-AP; Proteintech Technology, Wuhan, China), AKT (Cat. No. 4691; Cell Signaling Technology, Boston, MA, USA), phosphor-Akt (Ser473) (Cat. No. 4060; Cell Signaling Technology, Boston, MA, USA), Cleaved Caspase-3 (Cat. No. 9661; Cell Signaling Technology, Boston, MA, USA), Caspase-3 (Cat. No. 9662; Cell Signaling Technology, Boston, MA, USA) GAPDH (Cat. No. 5174; Cell Signaling Technology, Boston, MA, USA), Cyclin E1 (Cat. No. ET1612-16; HUABIO Technology, Hangzhou, China) and PPAP2A (PLPP1, which was used in immunohistochemistry experiments) (Cat. No. NBP1-59011; Novus Biologicals, Littleton, CO, USA) were used.

Techniques: Expressing, RNA Sequencing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Z-Ligustilide Combined with Cisplatin Reduces PLPP1-Mediated Phospholipid Synthesis to Impair Cisplatin Resistance in Lung Cancer

doi: 10.3390/ijms242317046

Figure Lengend Snippet: PLPP1 expression was low and associated with poor prognosis in lung cancer. ( A ) PLPP1 expression was detected in LUAD, LUSC, and corresponding normal tissues. LUAD (num (T) = 483; num (N) = 347). LUSC (num (T) = 486; num (N) = 338). * p < 0.05. ( B ) Representative images of the immunohistochemical staining of PLPP1 in lung cancer samples. Scale bars, 50 μm. −: negative expression; +: lower expression; ++: moderate expression; +++: high expression. The score criteria are described in detail in Materials and Methods. ( C ) The association between the PLPP1 protein expression and overall survival in patients with lung cancer. Low PLPP1 expression (n = 41), high PLPP1 expression (n = 35). * p < 0.05. ( D ) The correlation between PLPP1 expression and overall survival in 1451 lung cancer patients. ( E ) The correlation between PLPP1 expression and overall survival in 116 lung cancer patients administered chemotherapy. ( F ) The correlation between PLPP1 expression and overall survival in 268 lung cancer patients without chemotherapy.

Article Snippet: The following primary antibodies for BAX (Cat. No. 50599-2-Ig; Proteintech Technology, Wuhan, China), PLPP1 (PPAP2A, which was used in western blot experiments) (Cat. No. 17967-1-AP; Proteintech Technology, Wuhan, China) Cyclin A2 (Cat. No. 18202-1-AP; Proteintech Technology, Wuhan, China), AKT (Cat. No. 4691; Cell Signaling Technology, Boston, MA, USA), phosphor-Akt (Ser473) (Cat. No. 4060; Cell Signaling Technology, Boston, MA, USA), Cleaved Caspase-3 (Cat. No. 9661; Cell Signaling Technology, Boston, MA, USA), Caspase-3 (Cat. No. 9662; Cell Signaling Technology, Boston, MA, USA) GAPDH (Cat. No. 5174; Cell Signaling Technology, Boston, MA, USA), Cyclin E1 (Cat. No. ET1612-16; HUABIO Technology, Hangzhou, China) and PPAP2A (PLPP1, which was used in immunohistochemistry experiments) (Cat. No. NBP1-59011; Novus Biologicals, Littleton, CO, USA) were used.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: International Journal of Molecular Sciences

Article Title: Z-Ligustilide Combined with Cisplatin Reduces PLPP1-Mediated Phospholipid Synthesis to Impair Cisplatin Resistance in Lung Cancer

doi: 10.3390/ijms242317046

Figure Lengend Snippet: Z-ligustilide+cisplatin induced the reversal of resistance mediated by PLPP1. ( A ) Protein expression of PLPP1 in shCtrl and PLPP1 knockdown A549/DDP and H460/DDP cells, respectively. ( B ) Cell viability was measured in shCtrl and PLPP1 knockdown A549/DDP and H460/DDP cells with or without the Z-ligustilide and cisplatin combination treatment, *** p < 0.001. ( C ) Relative protein expressions in shCtrl and PLPP1 knockdown A549/DDP and H460/DDP cells with or without the Z-ligustilide and cisplatin combination treatment.

Article Snippet: The following primary antibodies for BAX (Cat. No. 50599-2-Ig; Proteintech Technology, Wuhan, China), PLPP1 (PPAP2A, which was used in western blot experiments) (Cat. No. 17967-1-AP; Proteintech Technology, Wuhan, China) Cyclin A2 (Cat. No. 18202-1-AP; Proteintech Technology, Wuhan, China), AKT (Cat. No. 4691; Cell Signaling Technology, Boston, MA, USA), phosphor-Akt (Ser473) (Cat. No. 4060; Cell Signaling Technology, Boston, MA, USA), Cleaved Caspase-3 (Cat. No. 9661; Cell Signaling Technology, Boston, MA, USA), Caspase-3 (Cat. No. 9662; Cell Signaling Technology, Boston, MA, USA) GAPDH (Cat. No. 5174; Cell Signaling Technology, Boston, MA, USA), Cyclin E1 (Cat. No. ET1612-16; HUABIO Technology, Hangzhou, China) and PPAP2A (PLPP1, which was used in immunohistochemistry experiments) (Cat. No. NBP1-59011; Novus Biologicals, Littleton, CO, USA) were used.

Techniques: Expressing, Knockdown

Journal: International Journal of Molecular Sciences

Article Title: Z-Ligustilide Combined with Cisplatin Reduces PLPP1-Mediated Phospholipid Synthesis to Impair Cisplatin Resistance in Lung Cancer

doi: 10.3390/ijms242317046

Figure Lengend Snippet: Z-ligustilide+cisplatin induced the inactivation of AKT by inhibiting PLPP1-mediated phospholipid synthesis. ( A ) The relative enrichments of each lipid class in shCtrl and PLPP1 knockdown A549/DDP cells with or without the Z-ligustilide and cisplatin combination treatment. ns, not significant, *** p < 0.001. ( B ) Protein expression of p-AKT in shCtrl and PLPP1 knockdown A549/DDP and H460/DDP cells with or without the Z-ligustilide and cisplatin combination treatment. ( C , D ) PI3K kinase activity assay and PIP 3 level measurements in shCtrl and PLPP1 knockdown A549/DDP and H460/DDP cells with or without the Z-ligustilide and cisplatin combination treatment. ns, not significant, ** p < 0.01 and *** p < 0.001.

Article Snippet: The following primary antibodies for BAX (Cat. No. 50599-2-Ig; Proteintech Technology, Wuhan, China), PLPP1 (PPAP2A, which was used in western blot experiments) (Cat. No. 17967-1-AP; Proteintech Technology, Wuhan, China) Cyclin A2 (Cat. No. 18202-1-AP; Proteintech Technology, Wuhan, China), AKT (Cat. No. 4691; Cell Signaling Technology, Boston, MA, USA), phosphor-Akt (Ser473) (Cat. No. 4060; Cell Signaling Technology, Boston, MA, USA), Cleaved Caspase-3 (Cat. No. 9661; Cell Signaling Technology, Boston, MA, USA), Caspase-3 (Cat. No. 9662; Cell Signaling Technology, Boston, MA, USA) GAPDH (Cat. No. 5174; Cell Signaling Technology, Boston, MA, USA), Cyclin E1 (Cat. No. ET1612-16; HUABIO Technology, Hangzhou, China) and PPAP2A (PLPP1, which was used in immunohistochemistry experiments) (Cat. No. NBP1-59011; Novus Biologicals, Littleton, CO, USA) were used.

Techniques: Knockdown, Expressing, Kinase Assay