Journal: Frontiers in Oncology

Article Title: TPL Inhibits the Invasion and Migration of Drug-Resistant Ovarian Cancer by Targeting the PI3K/AKT/NF-κB-Signaling Pathway to Inhibit the Polarization of M2 TAMs

doi: 10.3389/fonc.2021.704001

Figure Lengend Snippet: Triptolide (TPL) inhibition of the polarization induction of M2-type macrophages may be accomplished by inhibiting the PI3K/AKT/NF-κB pathway. (A) Images of tumors in the M, DDP, TPL, and DDP + TPL groups on day 15. (B) Weights (n = 3) of tumors in different treatment groups. (C) Kaplan–Meier survival curves of model mice bearing ovarian cancer after the intraperitoneal administration of TPL or DDP (n = 5). (D) Representative immunohistochemical expression of CD206 and CD31 in tumor tissues of different treatment groups. (E, F) Relative expression of MMP9, MMP2, VEGF, p-PI3K, PI3K, p-AKT, p-p65, AKT, and p65 in tumor tissues. *P < 0.05, **P < 0.01.

Article Snippet: The PVDF membranes were incubated with rabbit anti-MMP2 (1:1000; Proteintech, Cat. no.: 10373-2-AP), rabbit anti-Arg-1 (1:1000; CST, Cat. no.: 93668T), rabbit anti-MMP9 (1:1000; Proteintech, Cat. no.: 10375-2-AP), rabbit anti-VEGF (1:1000; Proteintech, Cat. no.: 19003-1-AP), rabbit anti-p65 (1:1000; Proteintech, Cat. no.: 10745-1-AP), rabbit anti-p-p65 (1:1000; CST, Cat. no.: 3033S), anti-AKT (1:1000; Proteintech, Cat. no.: 10176-2-AP), mouse anti-p-AKT (1:1000; Proteintech, Cat. no.: 66444-1-Ig), and mouse anti-β-actin (1:5000; Proteintech, Cat. no.: 60008-1-Ig) primary antibodies overnight at 4°C after blocking with 5% skim milk.

Techniques: Inhibition, Immunohistochemical staining, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Inhibition of PHLPP1 ameliorates cardiac dysfunction via activation of the PI3K/Akt/mTOR signalling pathway in diabetic cardiomyopathy

doi: 10.1111/jcmm.15123

Figure Lengend Snippet: Effects of PHLPP1 on myocardial apoptosis and fibrosis. A, Immunostaining of PHLPP1 (first row, n = 6) and cell apoptosis as determined by TUNEL assay (second‐fourth row, n = 5): apoptosis cell stained red; nuclei stained blue with DAPI. B, Cell apoptosis rate determined by TUNEL assay. C, The levels of cleaved caspase‐3 following PHLPP1 inhibition were measured by Western blot (n = 6). D, The levels of Bax and Bcl‐2 following PHLPP1 inhibition were measured by Western blot (n = 6). E, Representative Masson's trichrome staining (first row) and Sirius red staining (second and third rows) of the myocardium (n = 6). Immunostaining of collagen I (fourth row) and collagen III (fifth row) (n = 6). F, Quantification of Masson's trichrome staining (n = 6). G, Quantification of Sirius red staining (n = 6). H‐K, Western blot analysis of the protein expression of collagen I (F), collagen III (G), MMP2 (H) and MMP9 (I) (n = 6). Control: normal rats. DM: diabetes mellitus. shN.C: negative control shRNA. shPHLPP1: PHLPP1 shRNA. All experiments were performed at least 3 times. Data are expressed as the means ± SD. Statistical analysis was performed using one‐way ANOVA followed by Bonferroni's post hoc test. * P < .05 compared with control, and # P < .05 compared with DM or shN.C in DM

Article Snippet: The membranes were blocked in 5% non‐fat milk for 90 minutes at room temperature and then incubated at 4°C with specific primary antibodies against GAPDH (Abways, P04406), PHLPP1 (Proteintech, 22789‐1‐AP), collagenI (novusbio, NBP1‐30054), collagenIII (novusbio, NB600‐594SS), MMP2 (Proteintech, 10373‐2‐AP), MMP9 (Proteintech, 10387‐2‐AP), cleaved caspase‐3, Bcl2‐associated X protein (Bax) (CST, 2772), B‐cell lymphoma/leukaemia‐2 (Bcl‐2) (Affinity, AF6139), p‐PI3K (CST, 4228), p‐Akt (abcam, ab38449), p‐mTOR (abcam, ab109268), PI3K (CST, 4292), Akt (abcam, ab179463) and mTOR (abcam, ab134903) for detection.

Techniques: Immunostaining, TUNEL Assay, Staining, Inhibition, Western Blot, Expressing, Negative Control, shRNA