Journal: Materials Today Bio

Article Title: Matrix-bound EGF promotes malignant phenotypes of breast cancer organoids in the biomimetic ECM of alginate

doi: 10.1016/j.mtbio.2025.101818

Figure Lengend Snippet: Western blots of PI3K-AKT pathway showing expression of STAT6 and p-STAT6 (A), ERK1/2 and p-ERK1/2 (B), PI3K, p-PI3K, AKT, p-AKT, mTOR and p-mTOR (C) and ratios of the activated protein to the total protein were calculated. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, n = 3, analyzed by two-tailed Student's t-test.

Article Snippet: For Western Blot, anti-GAPDH (5174T), anti-ERK (4695T), anti- (phosphor) -ERK (9472T), anti-STAT (4904T) anti- (phosphor) -STAT (9145T), anti-AKT (9272T), anti- (phosphor) -AKT (4048T), anti-mTOR (2983T) and anti- (phosphor) - mTOR (5536T) were purchased from Cell Signaling Technology (USA), anti-E-cadherin (ab314063), anti-vimentin (ab20346), anti-PI3K (ab191606) and anti- (phosphor)-PI3K (ab278545) were purchased from Abcam (USA).

Techniques: Western Blot, Expressing, Two Tailed Test

Journal: World Journal of Gastroenterology

Article Title: Chaperonin-containing tailless complex polypeptide 1 subunit 6A negatively regulates autophagy and protects colorectal cancer cells from cisplatin-induced cytotoxicity

doi: 10.3748/wjg.v31.i18.105729

Figure Lengend Snippet: Chaperonin-containing tailless complex polypeptide 1 subunit 6A affects the autophagy pathway in colorectal cancer cells. A and B: Cell lysates were extracted from knockout (KO)-Control (Ctrl) and KO-chaperonin-containing tailless complex polypeptide 1 subunit 6A (CCT6A) HT29 cells, as well as overexpression (OE)-Ctrl and OE-CCT6A SW480 cells, and then subjected to immunoblotting using the indicated antibodies; C: Following treatment with chloroquine diphosphate salt (CQ, 20 μM, hereafter unless otherwise indicated) for 2 hours, immunoblotting was performed with the total proteins extracted from the indicated cells. DDP: Cisplatin; LC3: Light chain 3; mTOR: Mammalian target of rapamycin; Ulk1: Unc-51-like autophagy activating kinase 1.

Article Snippet: Primary antibodies against phosphorylated mammalian target of rapamycin (phosphor-mTOR) (S2448, 5536S), total-mTOR (2983S), phospho-p70 (T389, 9234S), total-p70 (34475S), total-mixed lineage kinase domain-like (MLKL, 14993S), total-RIP3 (13526S), phospho-RIP1 (S166, 65746S), phospho-RIP3 (S227, 93654S), Bcl-extra large (Bcl-xL) (2764S), poly(ADP-ribose) polymerase 1 (PARP-1, 9542S), and cleaved caspase-3 (9661S) were purchased from Cell Signaling Technology (Danvers, MA, United States).

Techniques: Knock-Out, Control, Over Expression, Western Blot

Journal: bioRxiv

Article Title: Lysosomal cathepsin D regulates bone turnover through distinct mode of actions of the autophagy pathways in osteoblasts and osteoclasts

doi: 10.1101/2025.04.09.645406

Figure Lengend Snippet: (A, B) PCR array shows that transcription profiles associated with the autophagy pathway are altered in MC3T3E1 osteoblastic cells with siRNA mediated knockdown of CtsD. Shown is the hierarchical clustering analysis of the PCR array data ( A ) and illustration of the mRNA fold-change of the top three upregulated and 21 downregulated autophagy pathway genes ( B ). (C) The knockdown of CtsD in MC3T3 cells was achieved by siRNA approach, and the cell lysates were immunoblotted with antibodies against phospho-PI3K p85, PI3K p85, phosphor-Akt, Akt, phospho-mTOR, and mTOR. Β-actin was used as internal control. (D - I) The densitometric quantification of blots for p-PI3K p85, PI3K p85, p-Akt, Akt, p-mTOR, and mTOR in (C) were shown in (D), (E), (F), (G), (H) and (I), respectively. * P < 0.05; ** P < 0.01. n = 3.

Article Snippet: The membrane was blocked with 5% non-fat dry milk and hybridized with antibodies against CtsD (Santa Cruz, CA, USA), LC3B, P62, phospho-Akt, Akt, phosphor-PI3K p85, PI3K p85, phosphor-mTOR, mTOR and β – actin (Cell Signaling, USA) at 4°C overnight.

Techniques: Knockdown, Control

Journal: bioRxiv

Article Title: Lysosomal cathepsin D regulates bone turnover through distinct mode of actions of the autophagy pathways in osteoblasts and osteoclasts

doi: 10.1101/2025.04.09.645406

Figure Lengend Snippet: (A, B) PCR array gene profiling shows that autophagy gene profiles are altered in RAW264.7 osteoclast precursor cells with siRNA mediated knockdown of CtsD. Hierarchical clustering analysis of the PCR array data ( A ) and illustration of the mRNA fold-change of the top 10 upregulated and 10 downregulated autophagy pathway genes ( B ). (C) The disruption of CtsD in RAW264.7 cells was performed with siRNA mediated knockdown, the cell lysates were immunoblotted with antibodies against p-PI3K p85, PI3K p85, p-Akt, Akt, p-mTOR, and mTOR. β-actin was used as internal control. (D - I) The densitometric quantification of the immunoblots for p-PI3K p85, PI3K p85, p-Akt, Akt, p-mTOR, and mTOR in (C). * P < 0.05; ** P < 0.01. n = 3.

Article Snippet: The membrane was blocked with 5% non-fat dry milk and hybridized with antibodies against CtsD (Santa Cruz, CA, USA), LC3B, P62, phospho-Akt, Akt, phosphor-PI3K p85, PI3K p85, phosphor-mTOR, mTOR and β – actin (Cell Signaling, USA) at 4°C overnight.

Techniques: Knockdown, Disruption, Control, Western Blot

Journal: Pharmaceuticals

Article Title: RNAi Screen Identifies AXL Inhibition Combined with Cannabinoid WIN55212-2 as a Potential Strategy for Cancer Treatment

doi: 10.3390/ph17111465

Figure Lengend Snippet: The anti-tumor activity of AXL inhibitor plus WIN55212-2 treatment in the MC38 cell xenograft mice. C57BL/6J mice were injected subcutaneously in the right flank with MC38 colon cancer cells at a density of 2.5 × 10 5 and treated with 50 mg/kg TP-0903 alone or in combination with WIN55212-2 (2.5 mg/kg, i.p.) every two days. CTX (80 mg/kg) was injected once as a control group. ( A ) The tumor weight of individual mice was detected at the end of the experiments. The infiltration of T lymphocytes in the tumor tissues was evaluated by flow cytometry. The percentages of CD3 + cells in CD45 + cells ( B ), CD8 + cells in CD3 + cells and CD4 + cells in CD3 + cells ( C ) infiltrating tumor sections were shown in the diagram. ( D ) The expressions of Ser473-phosphorylated AKT (p-AKT Ser473 ), Ser2448-phosphorylated mTOR (p-mTOR Ser2448 ), Tyr705-phosphorylated STAT3 (p-STAT3 Tyr705 ), AKT, mTOR and STAT3 in tumor tissues were evaluated by Western blot analysis. * p < 0.05, ** p < 0.01, *** p < 0.001 indicate a significant difference.

Article Snippet: Antibodies to phosphor-AXL Tyr698 , phosphor-AXL Tyr702 , AXL, phosphor-AKTser473, AKT, phosphor-mTOR Ser2448 , mTOR, phosphor-STAT3 Tyr705 , STAT3 and β-actin were purchased from Cell Signaling Technologies (Danvers, MA, USA).

Techniques: Activity Assay, Injection, Control, Flow Cytometry, Western Blot