migf1  (R&D Systems)

Journal: Oncology Letters

Article Title: Monocarboxylate transporter 4 promotes the migration of non‑cancerous L929 fibroblast cells by activating the IGF1/IGF1R/PIK3R3/SGK1 axis

doi: 10.3892/ol.2023.14047

Figure Lengend Snippet: IGF1 promotes the migration of control-L929 cells. (A) Representative wound healing images of the control-L929 and MCT4-L929 cells treated with or without SF and mIGF1 and (B) quantified wound healing results. (C) Western blot analysis of the expression levels of IGF1, IGF1R, PIK3R3 and SGK1 and the phosphorylated forms of IGF1R, PIK3R3 and SGK1 in control-L929 and MCT4-L929 cells supplemented with or without SF and mIGF1 using β-actin as a loading control and (D) semi-quantification of the results. The expression level of target genes in MCT4-L929 cells was normalized to that in control-L929 cells. (E) Phosphorylated/total protein ratio. The means of the data from three cell lines in each group are shown, and each error bar represents one standard deviation. Differences between groups were analyzed using two-tailed unpaired t-tests. *P<0.05, **P<0.01 and ***P<0.001. ns, not significant; IGF1, insulin-like growth factor 1; MCT4, monocarboxylate transporter 4; SF, serum-free starvation overnight before mIGF1 treatment; mIGF1, mouse IGF1; IGF1R, IGF1 receptor; PIK3R3, phosphoinositide 3-kinase regulatory subunit 3; SGK1, serum/glucocorticoid regulated kinase 1; p-, phosphorylated.

Article Snippet: Control-L929 and MCT4-L929 cells were seeded into 6-cm dishes and incubated in the presence or absence of recombinant mIGF1 (cat. no. 50437-MNAY; Sino Biological, Inc.) at a concentration of 500 pg/ml for 1 h or inhibitors (300 nM PPP/5 µM GSK650394) for 2 h in serum-free medium at 37°C.

Techniques: Migration, Western Blot, Expressing, Standard Deviation, Two Tailed Test

Journal: Oncology Letters

Article Title: Monocarboxylate transporter 4 promotes the migration of non‑cancerous L929 fibroblast cells by activating the IGF1/IGF1R/PIK3R3/SGK1 axis

doi: 10.3892/ol.2023.14047

Figure Lengend Snippet: Transcriptome data analysis. (A) Sample correlation analysis. The left and upper sides indicate sample clustering, while the right and lower sides of the figure are cell line names. Different colors of the squares represent the strength of the correlation between the two corresponding samples. (B) Bar chart showing the number of upregulated and downregulated DEGs. (C) Volcano plot of DEGs in which the x-axis represents log 2 |fold change| for MCT4-L929/control-L929 and the y-axis shows the log 10 (P-value). The two vertical dashed lines in the figure indicate the two-fold expression difference threshold, while the horizontal dashed line indicates the P=0.05 threshold. The red and green dots represent upregulated and downregulated genes, respectively. The gray dots represent genes that are not significantly and differentially expressed. (D) Heatmap of DEGs in which each column is a cell line and the y-axis represents DEGs. The colors indicate the expression levels of the DEGs; red and green represent upregulated and downregulated genes, respectively. (E) GO enrichment and (F) KEGG enrichment. The color of each bubble represents the P-value, while the size represents the number of DEGs in the GO or KEGG term. The x-axis shows the rich factor, which represents the ratio of DEGs vs. total genes in the pathway that is measured. DEGs, differentially expressed genes; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; Igf1, insulin-like growth factor 1; NoDiff, no difference.

Article Snippet: The culture supernatant was then collected to measure the IGF1 levels using a mouse IGF1 (mIGF1) ELISA kit (cat. no. EK0378; Boster Biological Technology), following the manufacturer's instructions.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Monocarboxylate transporter 4 promotes the migration of non‑cancerous L929 fibroblast cells by activating the IGF1/IGF1R/PIK3R3/SGK1 axis

doi: 10.3892/ol.2023.14047

Figure Lengend Snippet: MCT4 activates the IGF1/IGF1R/PIK3R3/SGK1 axis. (A) WB analysis of MCT4 and IGF1 levels in control-L929 and MCT4-L929 cell lines using β-actin as a loading control and (B) semi-quantification of the WB results. The expression of IGF1 was normalized to that of β-actin. (C) IGF1 concentration in the supernatant of MCT4-L929 and control-L929 cell lines as quantified using an enzyme-linked immunosorbent assay. (D) The mRNA levels of selected genes in the IGF1/IGF1R pathway in MCT4-L929 and control-L929 cells as quantified using reverse transcription-quantitative PCR. Transcription levels in MCT4-L929 cells were normalized to those in control-L929 cells. (E) WB analysis of IGF1, IGF1R, PIK3R3, AKT1/2/3 and SGK1 expression levels and the phosphorylation of IGF1R, PIK3R3, AKT1/2/3 and SGK1 and (F) semi-quantification of the WB results. (G) Phosphorylated/total protein ratio. The expression levels in MCT4-L929 cells were normalized to those in control-L929 cells. The means of the data from three cell lines in each group are shown, and each error bar represents one standard deviation. Differences between groups were analyzed using two-tailed unpaired t-tests. *P<0.05, **P<0.01 and ***P<0.001. ns, not significant; MCT4, monocarboxylate transporter 4; IGF1, insulin-like growth factor 1; IGF1R, IGF1 receptor; PIK3R3, phosphoinositide 3-kinase regulatory subunit 3; SGK1, serum/glucocorticoid regulated kinase 1; WB, western blotting; p-, phosphorylated.

Article Snippet: The culture supernatant was then collected to measure the IGF1 levels using a mouse IGF1 (mIGF1) ELISA kit (cat. no. EK0378; Boster Biological Technology), following the manufacturer's instructions.

Techniques: Expressing, Concentration Assay, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Standard Deviation, Two Tailed Test, Western Blot

Journal: Oncology Letters

Article Title: Monocarboxylate transporter 4 promotes the migration of non‑cancerous L929 fibroblast cells by activating the IGF1/IGF1R/PIK3R3/SGK1 axis

doi: 10.3892/ol.2023.14047

Figure Lengend Snippet: IGF1 promotes the migration of control-L929 cells. (A) Representative wound healing images of the control-L929 and MCT4-L929 cells treated with or without SF and mIGF1 and (B) quantified wound healing results. (C) Western blot analysis of the expression levels of IGF1, IGF1R, PIK3R3 and SGK1 and the phosphorylated forms of IGF1R, PIK3R3 and SGK1 in control-L929 and MCT4-L929 cells supplemented with or without SF and mIGF1 using β-actin as a loading control and (D) semi-quantification of the results. The expression level of target genes in MCT4-L929 cells was normalized to that in control-L929 cells. (E) Phosphorylated/total protein ratio. The means of the data from three cell lines in each group are shown, and each error bar represents one standard deviation. Differences between groups were analyzed using two-tailed unpaired t-tests. *P<0.05, **P<0.01 and ***P<0.001. ns, not significant; IGF1, insulin-like growth factor 1; MCT4, monocarboxylate transporter 4; SF, serum-free starvation overnight before mIGF1 treatment; mIGF1, mouse IGF1; IGF1R, IGF1 receptor; PIK3R3, phosphoinositide 3-kinase regulatory subunit 3; SGK1, serum/glucocorticoid regulated kinase 1; p-, phosphorylated.

Article Snippet: The culture supernatant was then collected to measure the IGF1 levels using a mouse IGF1 (mIGF1) ELISA kit (cat. no. EK0378; Boster Biological Technology), following the manufacturer's instructions.

Techniques: Migration, Western Blot, Expressing, Standard Deviation, Two Tailed Test

Journal: Oncology Letters

Article Title: Monocarboxylate transporter 4 promotes the migration of non‑cancerous L929 fibroblast cells by activating the IGF1/IGF1R/PIK3R3/SGK1 axis

doi: 10.3892/ol.2023.14047

Figure Lengend Snippet: A schematic illustration showing the mechanism in which MCT4 drives the migration of non-cancerous L929 fibroblast cells via activation of the IGF1/IGF1R/PIK3R3/SGK1 axis. MCT4, monocarboxylate transporter 4; IGF1, insulin-like growth factor 1; IGF1R, IGF1 receptor; PIK3R3, phosphoinositide 3-kinase regulatory subunit 3; SGK1, serum/glucocorticoid regulated kinase 1.

Article Snippet: The culture supernatant was then collected to measure the IGF1 levels using a mouse IGF1 (mIGF1) ELISA kit (cat. no. EK0378; Boster Biological Technology), following the manufacturer's instructions.

Techniques: Migration, Activation Assay