Fig 1: M2-EV-miR-365 facilitates the PDAC progression through the BTG2/FAK/AKT axis. (A) MiR-365 and BTG2 expression in PANC-1 and BxPC-3 cells measured by RT-qPCR. (B) Protein levels of BTG2 and FAK/AKT pathway-related proteins in PANC-1 and BxPC-3 cells examined by Western blot assay. (C) Proliferation of PANC-1 and BxPC-3 cells assessed by CCK-8. (D) Invasion of PANC-1 and BxPC-3 cells from different groups examined by Transwell (200×, scale bar = 50 µm). (E) Migration of PANC-1 and BxPC-3 cells from different groups was examined by Transwell (200×, scale bar = 50 µm). (F) Protein level of proliferation-related factors (Ki67, PCNA), apoptotic factors (Bax, Bcl-2, cleaved caspase-3) in different groups measured by Western blot assay. (G) Protein levels of EMT-related factors (Vimentin, Snail, E-cadherin) determined by Western blot assay. * in comparison to oe-NC, P < 0.05. # in comparison to M2-EVs + oe-NC, P < 0.05. NS, non-statistically significant. Measurement data were expressed as mean ± SD. Data between two groups were compared using unpaired t test. Comparisons among multiple groups were conducted by one-way ANOVA. Experiments were performed 3 times
Fig 2: The protein strip diagram and protein expressions level of PI3K-Akt pathway related proteins in each group. (A) Band diagrams of p-Akt, p-PI3K, Akt, and PI3K in brain tissue of each group of rats determined by western blot. (B) The expression of p-Akt in brain tissue of each group rats determined by western blot. (C) The expression of p-PI3K in brain tissue of each group rats determined by western blot. (D) The expression of Akt in brain tissue of each group rats determined by western blot. (E) The expression of PI3K in brain tissue of each group rats determined by western blot (n = 4). Compared with the sham operation group, ▲▲p < 0.01; compared with the MCAO group, *p < 0.05, **p < 0.01; compared with the LY294002 group, #p < 0.05, ##p < 0.01. Sham, sham operation group; MCAO, middle cerebral artery occlusion; LY294002, 2-(4-morpholinyl)-8-phenyl-chromone; DHI, Danhong Injection; NMDP, nimodipine.
Fig 3: Lnc‐ISG20 promotes fibrosis of MCs through miR‐486‐5p/NFAT5/p‐AKT. A, The mRNA levels of NFAT5 in MCs after treatment with oe‐lnc‐ISG20/oe‐NC + sh‐NC/sh‐NFAT5 determined by RT‐qPCR. B, The expression of NFAT5, p‐AKT, AKT in MCs after treatment with oe‐lnc‐ISG20/oe‐NC + sh‐NC/sh‐NFAT5 determined by RT‐qPCR. C, The protein expression of collagen IV, fibronectin and TGF‐β1 in MCs under unified HG treatment conditions after treatment with oe‐lnc‐ISG20/oe‐NC + sh‐NC/sh‐NFAT5 measured by Western blot assay. *P < .05 as compared to MCs treated with oe‐NC + sh‐NC. #P < .05 as compared to the MCs treated with oe‐lnc‐ISG20 + sh‐NC. (The data results are measurement data expressed by mean ± standard deviation. One‐way analysis of variance was used for comparisons among multiple groups followed by Tukey's post hoc test. The experiment was repeated 3 times)
Fig 4: Lnc‐ISG20 is highly expressed in DN patients, DN mouse kidney tissues and cell models. A, RT‐qPCR determination of the relative expression of lnc‐ISG20 in the normal kidney tissues of the control group (n = 30) and kidney tissues of DN patients (n = 30). B, Blood glucose levels in the control and DN mice. C, Urinary albumin excretion rate in the control and DN mice. D, HE staining showing pathological conditions of the kidney tissues in the control and DN mice. E, Masson staining displaying collagen fibres of kidney tissues in the control and DN mice. F, Immunohistochemistry for detection of protein expression of collagen IV, fibronectin and TGF‐β1 in the kidney tissues of the control and DN mice. G, RT‐qPCR determination of the relative expression of lnc‐ISG20 in kidney tissues of control mice (n = 10) and DN mice (n = 10). H, The expression of p‐AKT and total AKT proteins in kidney tissues of control mice (n = 10) and DN mice (n = 10) measured by Western blot assay. I, RT‐qPCR determination of the expression of lnc‐ISG20 in MCs under NG or HG condition. J, The protein expression of collagen IV, fibronectin and TGF‐β1 in MCs under NG or HG condition measured by Western blot assay. *P < .05 when compared with the normal tissues, the control mice or NG‐treated MCs (The results are measurement data expressed by mean ± standard deviation. An independent sample t test is used for the analysis between two groups, and the experiment is repeated 3 times)
Fig 5: FAK/AKT involves in the regulation of Kcnh2 in sepsis‐induced cardiac apoptosis. (A) The phosphorylation levels of AKT and FAK in Kcnh2+/‐ and WT rat cardiac extracts challenged with LPS (n = 3). (B) Immunohistostaining for phosphorylation status of AKT and FAK, (n = 3). Scale bar: 100 μm. (C) The phosphorylation levels of AKT and FAK in cardiomyocytes challenged by LPS at different time points (D) The phosphorylation change of AKT in NRCMs treated with bpV(HOpic).(n = 4). (E) TUNEL staining for heart treated with bpV(HOpic) and LPS, (n = 4), scale bar: 75 μm. (F) Western blot and quantitative analysis for the expression of FOXO3A, BCL‐2, BIM and PUMA in heart treated with bpV(HOpic) and LPS, (n = 5). (These results were performed as mean ± SD of at least 3 independent experiments. Statistical analysis was performed with one‐way ANOVA followed by Tukey's test. *, P < .05; **, P < .01; ***, P < .001 and ****, P < .0001)
Supplier Page from Abcam for Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798]