Journal: Journal of Functional Foods

Article Title: Phillygenin protects against osteoarthritis by repressing inflammation via PI3K/Akt/NF-κB signaling: In vitro and vivo studies

doi: 10.1016/j.jff.2021.104456

Figure Lengend Snippet: Fig. 3. The effect of PHI on the expression of IL-1β-induced matrix degrading enzymes in rat chondrocytes. Chondrocytes were stimulated with IL-1β in the presence of PHI or not for 24 h. (A, B) The expression of MMP13 and ADAMTS5 in chondrocytes and culture medium were measured by RT-PCR and ELISA. (C, D) Immu nofluorescence staining of MMP13 and type II collagen in chondrocytes (Scale Bar 50 μM). All values showed as mean values ± SD (##P < 0.01 vs control group. *P < 0.05, **P < 0.01 vs IL-1β group. n = 3).

Article Snippet: ELISA kits of PGE2, IL6, MMP13, TNF-α and ADAMTS5 were gained from R&D systems Inc. (MN, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Staining, Control

Journal: Shock (Augusta, Ga.)

Article Title: Restorative mechanisms regulating protein balance in skeletal muscle during recovery from sepsis

doi: 10.1097/SHK.0000000000000762

Figure Lengend Snippet: A-F: Representative Western blots and quantitation are presented for phosphorylated and/or total Akt, PRAS40, TSC2, AMPK, and REDD1. The final panel is a representative tubulin blot demonstrating equal protein loading. Loading controls (tubulin or actin) were run for all proteins examined, but only representative data are shown. Bar graphs: were quantified for phosphorylation of each protein normalized to the total amount of the respective protein, except for REDD1 where total protein was normalized to tubulin. Control values were set to 100 arbitrary units (AU). Values are mean ± SEM. *P < 0.05 sepsis-recovery compared to pair-fed controls values; n = 17 and 10, respectively.

Article Snippet: Antibodies included (Cell Signaling, Beverly, MA, unless otherwise noted): S6K1, S6K1 (Thr389), rpS6, rpS6 (Ser240/244 and Ser230/235), 4E-BP1 (Bethyl Laboratories, Montgomery, TX), 4E-BP1 (Ser65), eukaryotic elongation factor (eEF)2, eEF2 (Thr56), eEF2 kinase, eEF2K (Ser366; Dr. Chris Proud), ERK (extracellular signal-regulated kinses), ERK1/2 (Thr202/Tyr204), RSK (90 kDa ribosomal S6 kinase), RSK1/2 (Ser380), REDD1 (regulated in development and DNA damage responses; ProteinTech, Chicago, IL), Akt, Akt (Thr308 and Ser473), PRAS40 (proline-rich Akt substrate of 40 kDa), PRAS40 (Thr246), eIF4B, eIF4B (Ser422), AMP-activated protein kinase-α (AMPK), AMPKα (Thr172), tuberous sclerosis complex 2 (TSC2), TSC2 (Thr1462), Unc-51 like autophagy activating kinase 1 (ULK1), ULK1 (Ser757), p62 (aka SQSTM1), light-chain 3B (LC3B)-I and –II, and MyoD and myogenin.

Techniques: Western Blot, Quantitation Assay

Journal: Journal of Cancer

Article Title: Gambogic acid affects ESCC progression through regulation of PI3K/AKT/mTOR signal pathway

doi: 10.7150/jca.41115

Figure Lengend Snippet: Expression of apoptosis-related proteins and PI3K/AKT/mTOR pathway components in ESCC cells incubated with GA. (A) The expression levels of cleaved-PARP1, Bcl-2, Bax, cleaved-caspase 3 and cleaved-caspase9 were detected by using western blot analysis. (B) Western blotting was performed to analyze the levels of PI3K, p-AKT, p-mTOR, AKT, mTOR and PTEN.

Article Snippet: The following primary antibodies were applied in the experiments: cleaved-caspase3, cleaved-caspase9, cleaved-PARP1, phosphorylated AKT, Bcl-2, MMP2, MMP9 and β-actin (Proteintech Group, Wuhan, China); PARP1, BAX, PI3K, total AKT, phosphorylated AKT (ser473), p-mTOR (ser2448), cyclinB1 and cyclinD1 (Cell Signaling Technology, MA, USA). β-actin was chosen as a loading control in all experiments.

Techniques: Expressing, Incubation, Western Blot