Journal: Chinese Medicine

Article Title: Aidi injection inhibits the migration and invasion of gefitinib-resistant lung adenocarcinoma cells by regulating the PLAT/FAK/AKT pathway

doi: 10.1186/s13020-024-01054-1

Figure Lengend Snippet: The mechanism of action of PLAT in gefitinib-resistant NSCLC. A The mRNA expression of PLAT, ITGB3, TNC, FAK, AKT and PI3K after PLAT knockdown; B - C protein expression bands of t-PA, ITGB3, TNC, FAK, AKT, PI3K, p-FAK, p-AKT, p-PI3K, Vimentin, E-cadherin, N-cadherin and protein expression heatmap after PLAT knockdown; D the cell viability of PC9GR by PLAT knockdown; E the migration and invasion of PLAT knockdown (100 μm); F combination of PLAT (t-PA) with ITGB3 and TNC. * P < 0.05; ** P < 0.01

Article Snippet: The primary antibodies were as follows: t-PA polyclonal antibodies (1:800, Proteintech), CD61/integrin beta 3 polyclonal antibody (1:2500, Proteintech), TNC/Tenascin-C polyclonal antibodies (1:800, Proteintech), Vimentin polyclonal antibodies (1:800, Proteintech), pan-AKT/1/2/3 antibody (1:1000, Affinity), phospho-pan-AKT1/2/3 (Thr308) antibody (1:1000, Affinity), PI3K p85 alpha antibody (1:1000, Affinity), phospho-PI3K p85 alpha (Tyr607) antibody (1:1000, Affinity), FAK antibody (1:1000, Affinity), phospho-FAK (Tyr397) antibody (1:1000, Affinity), E-cadherin antibody (1:1000, Affinity), N-cadherin antibody (1:1000, Affinity), GAPDH polyclonal antibodies (1:20000, Proteintech).

Techniques: Expressing, Knockdown, Migration

Journal: Chinese Medicine

Article Title: Aidi injection inhibits the migration and invasion of gefitinib-resistant lung adenocarcinoma cells by regulating the PLAT/FAK/AKT pathway

doi: 10.1186/s13020-024-01054-1

Figure Lengend Snippet: The mRNA and protein expression in PC9 and PC9GR cells and the changes of mRNA and protein expression in PC9GR cells after different drug treatments. A mRNA expression levels of PLAT, ITGB3, TNC, FAK, AKT and PI3K in PC9 and PC9GR cells; B the effect of ADI on the mRNA expression of PLAT, ITGB3, TNC, FAK and PI3K in PC9GR cells; C the effect of ADI combined with gefitinib on the mRNA expression of PLAT, ITGB3, TNC, FAK, AKT and, PI3K in PC9GR cells; D bands of PLAT (t-PA), ITGB3, TNC, FAK, AKT, PI3K, p-FAK, p-AKT, p-PI3K, Vimentin, E-cadherin, N-cadherin in PC9 and PC9GR cells; E the protein expression of PLAT (t-PA), ITGB3, TNC, FAK, AKT, PI3K, p-FAK, p-AKT, p-PI3K, Vimentin, E-cadherin, N-cadherin in PC9 and PC9GR cells. * P < 0.05; ** P < 0.01

Article Snippet: The primary antibodies were as follows: t-PA polyclonal antibodies (1:800, Proteintech), CD61/integrin beta 3 polyclonal antibody (1:2500, Proteintech), TNC/Tenascin-C polyclonal antibodies (1:800, Proteintech), Vimentin polyclonal antibodies (1:800, Proteintech), pan-AKT/1/2/3 antibody (1:1000, Affinity), phospho-pan-AKT1/2/3 (Thr308) antibody (1:1000, Affinity), PI3K p85 alpha antibody (1:1000, Affinity), phospho-PI3K p85 alpha (Tyr607) antibody (1:1000, Affinity), FAK antibody (1:1000, Affinity), phospho-FAK (Tyr397) antibody (1:1000, Affinity), E-cadherin antibody (1:1000, Affinity), N-cadherin antibody (1:1000, Affinity), GAPDH polyclonal antibodies (1:20000, Proteintech).

Techniques: Expressing

Journal: Chinese Medicine

Article Title: Aidi injection inhibits the migration and invasion of gefitinib-resistant lung adenocarcinoma cells by regulating the PLAT/FAK/AKT pathway

doi: 10.1186/s13020-024-01054-1

Figure Lengend Snippet: The influence of Aidi injection and its combination with gefitinib on crucial genes and proteins. A The protein expression of PLAT (t-PA), ITGB3, TNC, FAK, AKT, PI3K, p-FAK, p-AKT, p-PI3K, Vimentin, E-cadherin, and N-cadherin in PC9GR after ADI treatment; B heatmap of protein expression of PLAT (t-PA), ITGB3, TNC, FAK, AKT, PI3K, p-FAK, p-AKT, p-PI3K, Vimentin, E-cadherin, and N-cadherin in PC9GR cells; C the protein expression of PLAT (t-PA), ITGB3, TNC, FAK, AKT, PI3K, p-FAK, p-AKT, p-PI3K, Vimentin, E-cadherin, and N-cadherin in PC9GR cells after ADI, gefitinib and ADI combined with gefitinib treatment; D expression heatmap of PLAT (t-PA), ITGB3, TNC, FAK, AKT, PI3K, p-FAK, p-AKT, p-PI3K, Vimentin, E-cadherin, and N-cadherin proteins in PC9GR cells after ADI, gefitinib and ADI combined with gefitinib treatment

Article Snippet: The primary antibodies were as follows: t-PA polyclonal antibodies (1:800, Proteintech), CD61/integrin beta 3 polyclonal antibody (1:2500, Proteintech), TNC/Tenascin-C polyclonal antibodies (1:800, Proteintech), Vimentin polyclonal antibodies (1:800, Proteintech), pan-AKT/1/2/3 antibody (1:1000, Affinity), phospho-pan-AKT1/2/3 (Thr308) antibody (1:1000, Affinity), PI3K p85 alpha antibody (1:1000, Affinity), phospho-PI3K p85 alpha (Tyr607) antibody (1:1000, Affinity), FAK antibody (1:1000, Affinity), phospho-FAK (Tyr397) antibody (1:1000, Affinity), E-cadherin antibody (1:1000, Affinity), N-cadherin antibody (1:1000, Affinity), GAPDH polyclonal antibodies (1:20000, Proteintech).

Techniques: Injection, Expressing

Journal: Cell Biology and Toxicology

Article Title: S100A2 activation promotes interstitial fibrosis in kidneys by FoxO1-mediated epithelial-mesenchymal transition

doi: 10.1007/s10565-024-09929-7

Figure Lengend Snippet: FoxO1 is a potential target of S100A2. ( A ) GO and ( B ) KEGG enrichment analyses for differentially expressed genes in TGF-β1-stimulated HK-2 cells with S100A2 knockdown. BP, CC, MF denote biological processes, cellular components, and molecular functions, respectively. S100A2 knockdown blocks ( C , F ) PI3K and activates ( D , G ) FoxO1. ( E , H ) Overexpression of S100A2 inhibits FoxO1 levels. ( I ) Co-IP demonstrates the interaction between S100A2 and FoxO1. ( J ) IF confirms the co-localization of S100A2 and FoxO1. Scale bar, 25 μm. ( K - M ) Immunoblot analysis of nuclear and cytoplasmic levels of p-FoxO1, FoxO1, p-Smad2/3, Snail1, and S100A2 in TGF-β1-stimulated HK-2 cells. ( N ) The effect of TGF-β1 treatment on the fluorescence intensity of p-FoxO1 in HK2 cells. * P < 0.05, ** P < 0.01, *** P < 0.001. ns, no statistical difference

Article Snippet: After blocking with 5% non-fat milk, the membrane was probed with the relevant primary antibodies overnight including anti-Vimentin (Santa Cruz), E-cadherin (Abcam), α-SMA (Santa Cruz), N-cadherin (Abcam), COL1 (Proteintech), S100A2 (Abcam), Smad2/3 (Abcam), p-Smad2/3 (Thr8, Abcam), p-PI3K (Tyr607, Affinity, USA), PI3K (Proteintech), p-AKT (Ser473, CST), AKT (CST), Sirt1 (Affinity), p-FoxO1 (Ser256, Affinity), FoxO1 (Affinity), Snail1 (Abcam), COL3 (Proteintech), GAPDH (Proteintech), H3 (Proteintech), and β-actin (Affinity).

Techniques: Knockdown, Over Expression, Co-Immunoprecipitation Assay, Western Blot, Fluorescence

Journal: Frontiers in Pharmacology

Article Title: Epimedin C Alleviates Glucocorticoid-Induced Suppression of Osteogenic Differentiation by Modulating PI3K/AKT/RUNX2 Signaling Pathway

doi: 10.3389/fphar.2022.894832

Figure Lengend Snippet: Epimedin C enhanced the OSX, RUNX2, and ALPL protein expression in MC3T3-E1 Cells treated with DEX. (A) Following 7 days of MC3T3-E1 cells treatment using epimedin C and DEX in OM, the protein was extracted by 10 percent SDS-PAGE and identified using the specified antibodies of OSX, RUNX2, and ALPL. As a loading control, β-actin was employed. (B) The bar charts depicted ImageJ’s measurements of RUNX2, OSX, and ALPL. DEX: dexamethasone; OM: osteogenic induction medium. * p < 0.05, ** p < 0.01.

Article Snippet: The following antibodies were utilized for incubation: anti-OSX (1:1000, ab209484, Abcam), Anti-ALPL (1:2000, ab65834, Abcam), Phospho-Akt (Ser473) (1:2000, #4511, Cell Signal Technology), anti-RUNX2 (1:1000, ab236639, Abcam), Akt (pan) (C67E7) (1:2000, #4691, Cell Signal Technology), Phospho-PI3K p85 alpha (Tyr607) (1:1000, #AF3241, Affinity), PI3 kinase P110 alpha Antibody (1:1000, #AF5112, Affinity) and anti-β-actin antibody (1:1000, AF0003, Beyotime).

Techniques: Expressing, SDS Page

Journal: Frontiers in Pharmacology

Article Title: Epimedin C Alleviates Glucocorticoid-Induced Suppression of Osteogenic Differentiation by Modulating PI3K/AKT/RUNX2 Signaling Pathway

doi: 10.3389/fphar.2022.894832

Figure Lengend Snippet: Epimedin C activated AKT and PI3K phosphorylation in MC3T3-E1 cells treated using DEX. (A) Western blotting was employed to assess the proportions of phosphorylated PI3K and AKT. As an endogenous control, β-actin was employed. (B,C) : ImageJ was used to quantify the p-PI3K/PI3K and p-AKT/AKT ratios as depicted by the bar charts. DEX: dexamethasone, OM: osteogenic induction medium. * p < 0.05, ** p < 0.01.

Article Snippet: The following antibodies were utilized for incubation: anti-OSX (1:1000, ab209484, Abcam), Anti-ALPL (1:2000, ab65834, Abcam), Phospho-Akt (Ser473) (1:2000, #4511, Cell Signal Technology), anti-RUNX2 (1:1000, ab236639, Abcam), Akt (pan) (C67E7) (1:2000, #4691, Cell Signal Technology), Phospho-PI3K p85 alpha (Tyr607) (1:1000, #AF3241, Affinity), PI3 kinase P110 alpha Antibody (1:1000, #AF5112, Affinity) and anti-β-actin antibody (1:1000, AF0003, Beyotime).

Techniques: Western Blot