Journal: Cell Communication and Signaling : CCS

Article Title: MIR4726 EccDNA drives bortezomib resistance in multiple myeloma by enhancing MIR4726-5p/NXF1/NKIRAS2 axis dependent autophagy

doi: 10.1186/s12964-025-02340-7

Figure Lengend Snippet: Downregulation of NKIRAS2 induced activation of the NF-κB pathway and increased autophagy. A LC3I, LC3II, p-p65, p65, and NKIRAS2 expression by WB in negative control or siNKIRAS2-transfected RPMI 8226 cells. B After treatment with 10 nmol/L bortezomib for 24 h, the autophagosomes in the RPMI 8226 WT and siNKIRAS2 cells were analyzed via TEM. C LC3I, LC3II, p-p65, p65, and NKIRAS2 expression in siNXF1-, OE-NKIRAS2 plasmids- or negative control-transfected RPMI 8226 cells as determined by WB. D After treatment with 10 nmol/L bortezomib for 24 h, TEM analysis of autophagosomes in siNXF1-, OE-NKIRAS2- or negative control-transfected RPMI 8226 cells was performed. NC: Negative control. Cell nucleus (N); Nucleolus (Nu); Mitochondria (M); Rough endoplasmic reticulum (RER); Lipid droplets (LD); Lysosomes (Ly); Autophagic lysosomes (↑); Autophagosome (↑); Golgi apparatus (Go)

Article Snippet: The paraffin slices were dewaxed to water, heated for 30 min with ethylene diamine tetra-acetic acid buffer (pH 8.0) (Servicebio, G1206), blocked with endogenous peroxidase, blocked with serum, and then incubated overnight at 4 °C in a wet box with the following primary antibodies: anti-LC3A/B rabbit pAb (Servicebio, GB11124, 1:200); anti-cleaved-caspase-3 rabbit pAb (Servicebio, GB11532-100, 1:500); anti-Ki67 rabbit pAb (Servicebio, GB111499 , 1:1000); and anti-phospho-NF-kB p65 rabbit pAb (Servicebio, GB113882 , 1:200).

Techniques: Activation Assay, Expressing, Negative Control, Transfection

Journal: Cell Communication and Signaling : CCS

Article Title: MIR4726 EccDNA drives bortezomib resistance in multiple myeloma by enhancing MIR4726-5p/NXF1/NKIRAS2 axis dependent autophagy

doi: 10.1186/s12964-025-02340-7

Figure Lengend Snippet: AgoMIR4726-5p promotes MM progression and reduces the antitumor effect of bortezomib in vivo. A Flow chart for the in vivo experiment (created with bioRender.com, with permission). B Tumor volume was calculated with the formula (length × width 2 )/2, and length and width were measured with electronic calipers. The frequency of measurements was adjusted according to the tumor growth rate. C Weights of the tumors at the end of the study. D Photographs of all mouse tumors in the four groups. E Representative immunohistochemical analysis of Ki67, LC3B, p-p65, and cleaved caspase 3 in tumors from the 4 groups of mice

Article Snippet: The paraffin slices were dewaxed to water, heated for 30 min with ethylene diamine tetra-acetic acid buffer (pH 8.0) (Servicebio, G1206), blocked with endogenous peroxidase, blocked with serum, and then incubated overnight at 4 °C in a wet box with the following primary antibodies: anti-LC3A/B rabbit pAb (Servicebio, GB11124, 1:200); anti-cleaved-caspase-3 rabbit pAb (Servicebio, GB11532-100, 1:500); anti-Ki67 rabbit pAb (Servicebio, GB111499 , 1:1000); and anti-phospho-NF-kB p65 rabbit pAb (Servicebio, GB113882 , 1:200).

Techniques: In Vivo, Immunohistochemical staining

Journal: bioRxiv

Article Title: Pharmacological inhibition of CXCR4 increases the anti-tumor activity of conventional and targeted therapies in B-cell lymphoma models

doi: 10.1101/2025.07.10.664047

Figure Lengend Snippet: REC1 cells were exposed to DMSO (control), SPX5551 (4 µM), ibrutinib (4 nM), or a combination of the two for 6 hours. Representative images of two independent experiments and protein quantification of RELA (cytoplasmatic, A) or RELB (nuclear, B). DAPI is stained blue, and RELA/RELB is stained red. Barplots on the left show the median fluorescence intensity, * for p<0.05, ** for p<0.01 from a Mann-Whitney U-test.

Article Snippet: It was followed by the incubation of samples at 4°C with primary rabbit monoclonal anti-human NF-kB p65 antibody (D14E12) (1/100; Cell Signaling) overnight.

Techniques: Control, Staining, Fluorescence, MANN-WHITNEY

Journal: Journal of Molecular and Cellular Cardiology Plus

Article Title: Independent and synergistic roles of MEK-ERK1/2 and PKC pathways in regulating functional changes in vascular tissue following flow cessation

doi: 10.1016/j.jmccpl.2025.100300

Figure Lengend Snippet: Effects of the MEK inhibitor, Trametinib, and PKC inhibitor, RO-317549, on ERK and NF-κB phosphorylation. BAs were incubated in OC for 6 h. (a) Western blot analysis of total ERK and p-ERK, (b) Quantitative data of ERK, (c) Western blot of total NF-κB and pNF-κB (d) Quantitative data of NF-κB Data are shown as mean ± SEM (Biological replicate; n = 5) and protein expression was compared by one-way ANOVA, with Dunnett's post-test. * = p < 0.05. Please note that the order of the inhibitor is not the same across the figures, due to the original experimental setup.

Article Snippet: The membranes were then transferred to a rocking table in room temperature with Blocking Buffer (EveryBlot Blocking Buffer, Cat. #: 12010020, Bio-Rad) for 10 min and then, the membranes were incubated with primary antibody dilutions overnight in 4 °C (1:1000 Mouse Anti-p44 / 42 MAP Kinase Monoclonal Antibody, Unconjugated, Clone L34F12 [Cell Signaling Technology Cat# 4696, lot: 29, RRID: AB_390780 ], 1:1000 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb [Cell Signaling Technology Cat# 4370, Lot: 28, RRID: AB_2315112 ), 1:1000 NF-κB p65 (L8F6) Mouse mAb [Cell Signaling Technology Cat# 6956, Lot:10, RRID: AB_10828935 ], 1:1000 Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb [Cell Signaling Technology Cat# 3033,Lot: 19, RRID: AB_331284 ], GAPDH (D16H11) XP Rabbit mAb, [Cell Signaling Technology Cat# 5174, Lot: 8, RRID: AB_10622025 ], 1:1000 Actin, Smooth Muscle (1A4) Mouse Monoclonal Antibody [Cell Marque Cat# 202 M-94, Lot:0000243213, RRID: AB_1157937 ], 1:1000 alpha smooth muscle Actin antibody (EPR5368) [Abcam Cat# ab124964, Lot: GR303485 –26, RRID: AB_11129103 ]).

Techniques: Phospho-proteomics, Incubation, Western Blot, Expressing

Journal: Journal of Molecular and Cellular Cardiology Plus

Article Title: Independent and synergistic roles of MEK-ERK1/2 and PKC pathways in regulating functional changes in vascular tissue following flow cessation

doi: 10.1016/j.jmccpl.2025.100300

Figure Lengend Snippet: The effect of NF-κB inhibitors in the contractility of BAs incubated for 48 h in OC. (a) Log concentration-response curves in response S6c, following incubation with the NF-κB inhibitors SP100030, BMS 345541, IMD 0354 (10 −6 M) the E max was compared by a multiple t -test with a Holm-Sidak correction (b) Log-concentration-response curves in response to S6c following incubation with 10 −5 M and 10 −6 M BMS 345541, the E max was compared by a multiple t-test with a Holm-Sidak correction (c) E max(S6c) contractions of RO317549 10 −6 M, BMS 345541 10 −6 M and their combination, compared with Trametinib 10 −8 M and its combination with BMS 345541 10 −6 M. E max were compared by one-way ANOVA, with Dunnett's post-test. Data are shown as mean ± SEM (Biological replicate; n = 4–18). * = p < 0.05.

Article Snippet: The membranes were then transferred to a rocking table in room temperature with Blocking Buffer (EveryBlot Blocking Buffer, Cat. #: 12010020, Bio-Rad) for 10 min and then, the membranes were incubated with primary antibody dilutions overnight in 4 °C (1:1000 Mouse Anti-p44 / 42 MAP Kinase Monoclonal Antibody, Unconjugated, Clone L34F12 [Cell Signaling Technology Cat# 4696, lot: 29, RRID: AB_390780 ], 1:1000 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb [Cell Signaling Technology Cat# 4370, Lot: 28, RRID: AB_2315112 ), 1:1000 NF-κB p65 (L8F6) Mouse mAb [Cell Signaling Technology Cat# 6956, Lot:10, RRID: AB_10828935 ], 1:1000 Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb [Cell Signaling Technology Cat# 3033,Lot: 19, RRID: AB_331284 ], GAPDH (D16H11) XP Rabbit mAb, [Cell Signaling Technology Cat# 5174, Lot: 8, RRID: AB_10622025 ], 1:1000 Actin, Smooth Muscle (1A4) Mouse Monoclonal Antibody [Cell Marque Cat# 202 M-94, Lot:0000243213, RRID: AB_1157937 ], 1:1000 alpha smooth muscle Actin antibody (EPR5368) [Abcam Cat# ab124964, Lot: GR303485 –26, RRID: AB_11129103 ]).

Techniques: Incubation, Concentration Assay

Journal: Journal of Molecular and Cellular Cardiology Plus

Article Title: Independent and synergistic roles of MEK-ERK1/2 and PKC pathways in regulating functional changes in vascular tissue following flow cessation

doi: 10.1016/j.jmccpl.2025.100300

Figure Lengend Snippet: Signaling pathways involved in ET B receptor upregulation and VSMC contraction. Extracellular stimuli such as flow cessation activate the MAPK/ERK pathway, which is disrupted by MEK inhibition (Trametinib) or ERK inhibition (Ulixertinib). In parallel, intracellular Ca 2+ increase leads to delayed PKC activation, promoting NF-κB signaling via the IKK complex. PKC and NF-κB inhibition (RO-317549 and BMS345541, respectively) remain effective when applied up to 6 h post-stimulus. Combined inhibition further suppresses ET B upregulation on VSMCs and reduces receptor-mediated contraction, indicating that both pathways contribute independently and are required for full functional response . Abbreviations: ET B : endothelin type B receptor; VSMC: vascular smooth muscle cell; NF-κB: nuclear factor. Created in BioRender. Kazantzi, S. (2025) https://BioRender.com/u89c445

Article Snippet: The membranes were then transferred to a rocking table in room temperature with Blocking Buffer (EveryBlot Blocking Buffer, Cat. #: 12010020, Bio-Rad) for 10 min and then, the membranes were incubated with primary antibody dilutions overnight in 4 °C (1:1000 Mouse Anti-p44 / 42 MAP Kinase Monoclonal Antibody, Unconjugated, Clone L34F12 [Cell Signaling Technology Cat# 4696, lot: 29, RRID: AB_390780 ], 1:1000 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb [Cell Signaling Technology Cat# 4370, Lot: 28, RRID: AB_2315112 ), 1:1000 NF-κB p65 (L8F6) Mouse mAb [Cell Signaling Technology Cat# 6956, Lot:10, RRID: AB_10828935 ], 1:1000 Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb [Cell Signaling Technology Cat# 3033,Lot: 19, RRID: AB_331284 ], GAPDH (D16H11) XP Rabbit mAb, [Cell Signaling Technology Cat# 5174, Lot: 8, RRID: AB_10622025 ], 1:1000 Actin, Smooth Muscle (1A4) Mouse Monoclonal Antibody [Cell Marque Cat# 202 M-94, Lot:0000243213, RRID: AB_1157937 ], 1:1000 alpha smooth muscle Actin antibody (EPR5368) [Abcam Cat# ab124964, Lot: GR303485 –26, RRID: AB_11129103 ]).

Techniques: Protein-Protein interactions, Inhibition, Activation Assay, Functional Assay