Journal: Cancer Research

Article Title: Human Rhomboid Family-1 Suppresses Oxygen-Independent Degradation of Hypoxia-Inducible Factor-1α in Breast Cancer

doi: 10.1158/0008-5472.can-13-1027

Figure Lengend Snippet: Figure 3. Inhibitory effect of RHBDF1 gene silencing in MCF7 cells on HIF1a stability and VEGF secretion. A, effect of RHBDF1 shRNA (shRHB) or scrambled shRNA (shScr) on HIF1a degradation in MCF7 cells in hypoxia. B, effect of shRHB or shScr on HIF1a protein stability in MCF7 cells during reoxygenation. C, nuclear translocation of HIF1a in shSrc- or shRHB-treated MCF7 cells; a-tubulin and Myc are cytoplasmic and nuclear protein controls, respectively. D, changes of VEGF concentrations in the condition media of shRHB (white bars)- or shScr (black bars)-treated MCF7 cells under normoxic or hypoxic conditions as a function of time (hours). E, changes of VEGF concentrations in the condition media of shRHB (white bars)- or shScr (black bars)-treated MDA-MB-231 cells under normoxic or hypoxic conditions as a function of time (hours). Each experiment was repeated two times. , P < 0.05, Student t test.

Article Snippet: Four independent shRNA 29-mers against RHBDF1 and a scrambled shRNA sequence were purchased from OriGene Technologies (cat# TF302009).

Techniques: shRNA, Translocation Assay

Journal: Cancer Research

Article Title: Human Rhomboid Family-1 Suppresses Oxygen-Independent Degradation of Hypoxia-Inducible Factor-1α in Breast Cancer

doi: 10.1158/0008-5472.can-13-1027

Figure Lengend Snippet: Figure 4. Involvement of RHBDF1, RACK1, and HSP90 in oxygen-independent HIF1a degradation. A, effect of RHBDF1 gene silencing in MCF7 cells on the stability of HA-tagged P402A/P564A mutant HIF1a (HA-HIF1a) in hypoxia. B, effect of RHBDF1 overexpression in T47D cells on the stability of the mutant HIF1a in hypoxia. C, effect of RACK1 gene silencing on HIF1a stability in MCF7 cells in hypoxia. D, effect of FLAG-RHBDF1 overexpression in T47D cells on the stability of the mutant HIF1a in hypoxia. E, effect of 17-AAG treatment (0.5 mmol/L, 24 hours) of RHBDF1 gene-silenced MCF7 cells on HIF1a stability in hypoxia. F, effect of 17-AAG treatment (0.5 mmol/L, 24 hours) of RHBDF1-overexpressing T47D cells on HIF1a in hypoxia. G, co-IP of RHBDF1 and RACK1 from MCF7 cells transfected with FLAG-RHBDF1. H, co-IP of RHBDF1 and HIF1a from MCF7 cells cotransfected with HA-tagged wild-type HIF1a. I, co-IP of RHBDF1 with HIF1a and RACK1 from MCF7 cells in the absence or presence of MG132 (10 mmol/L, 4 hours) in normoxia or hypoxia. J, co-IP of HIF1a with HSP90 and of RHBDF1 with RACK1 from MCF7 cells cotransfected with FLAG-RHBDF1 and the mutant HIF1a upon treatment with either control siScr, RACK1 siRNA (siRACK1), or 17-AAG (0.5 mmol/L). Each experiment was repeated two times and representative Western blotting analysis or co-IP results from one experiment are shown.

Article Snippet: Four independent shRNA 29-mers against RHBDF1 and a scrambled shRNA sequence were purchased from OriGene Technologies (cat# TF302009).

Techniques: Mutagenesis, Over Expression, Co-Immunoprecipitation Assay, Transfection, Control, Western Blot

Journal: Cancer Cell International

Article Title: Differential drug resistance acquisition to doxorubicin and paclitaxel in breast cancer cells

doi: 10.1186/s12935-014-0142-4

Figure Lengend Snippet: Impact of PXR activation in DOX and PTX treated cells. (A) PXR mRNA expression with the treatment of DOX and PTX at 0, 4, 8, 12, 24, 36, 48 h. (B) Western blot analysis of PXR with 24 h treatment of DOX and PTX. **, p < 0.01 (n = 3). (C) RT-PCR analysis of PXR mRNA in PTX only (PTX) or treated cells exposed to transfection conditions in the absence of siRNA (mock) or cells transfected with non-targeting (NC) or PXR-specific siRNA (siRNA). (D) Comparison of MDR1 mRNA expression in the cells exposed to transfection conditions with 8 h/36 h DOX and PTX treatment. **, p < 0.01 (n = 3). (E) Rho-123 fluorescence ratio in the cells exposed to transfection conditions with 8 h/36 h DOX and PTX treatment. *, p < 0.05 (n = 3).

Article Snippet: The siRNA sequence targeting human PXR gene and the negative control that consisted of a non-complementary sequence were developed by GenePharma (Shanghai, China).

Techniques: Activation Assay, Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Transfection, Comparison, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Apobec-1 Complementation Factor (A1CF) Inhibits Epithelial-Mesenchymal Transition and Migration of Normal Rat Kidney Proximal Tubular Epithelial Cells

doi: 10.3390/ijms17020197

Figure Lengend Snippet: Apobec-1 complementation factor (A1CF) is highly conserved among species. ( a ) A1CF amino acid sequences of human , mouse , rat , chicken , zebrafish , xenopus laevis were selected to analysis. The identical amino acids were shaded in black and the similar amino acids in grey; ( b ) Phylogenetic tree of A1CF amino acids is constructed using MEGA6 software, bootstrap percentages based on 1000 replicates, are shown on each branch. Identity values are listed on the right ranging from 78% to 92%, revealing that A1CF is highly conserved among species.

Article Snippet: SiRNA oligonucleotides were purchased from Shanghai GenePharma Co., Ltd. Two stealth siRNAs targeting different sequences of A1CF mRNA were selected to silence A1CF of Rattus norvegicus .

Techniques: Construct, Software

Journal: International Journal of Molecular Sciences

Article Title: Apobec-1 Complementation Factor (A1CF) Inhibits Epithelial-Mesenchymal Transition and Migration of Normal Rat Kidney Proximal Tubular Epithelial Cells

doi: 10.3390/ijms17020197

Figure Lengend Snippet: A1CF is expressed in mouse kidney starting at E15.5d and centers on the kidney tubules in C57BL/6 mice . ( a – c ) Whole mount in situ hybridization demonstrates that A1CF is expressed in the later stage of kidney development of C57BL/6 mice . White dot line indicates the area of metanephros rudiment (original magnification, ×40); ( c ′ ) To clearly prove the expression of A1CF, metanephros of embryonic day 15.5 (E15.5d) was detected with original magnification, ×100; ( d – g ) Section in situ hybridization demonstrates that in the adult mice kidney, A1CF is mainly expressed in the tubules of inner cortex but not in the other parts of the kidney section; ( d ) The black dot line marks the boundary between cortex and medulla. Scale bar represents 30 µm; ( e – g ) Images is the amplification of ( d ). ( e ) Scale bar represents 50 µm; ( f ) Image represents the field surrounding kidney tubules; Scale bar represents 100 µm; ( g ) Image represents the field of kidney tubules. Scale bar represents 100 µm; ( h , i ) In control groups no hybridization signal can be detected; ( h ) Scale bar represents 50 µm; ( i ) Scale bar represents 100 µm.

Article Snippet: SiRNA oligonucleotides were purchased from Shanghai GenePharma Co., Ltd. Two stealth siRNAs targeting different sequences of A1CF mRNA were selected to silence A1CF of Rattus norvegicus .

Techniques: In Situ Hybridization, Expressing, Amplification, Hybridization

Journal: International Journal of Molecular Sciences

Article Title: Apobec-1 Complementation Factor (A1CF) Inhibits Epithelial-Mesenchymal Transition and Migration of Normal Rat Kidney Proximal Tubular Epithelial Cells

doi: 10.3390/ijms17020197

Figure Lengend Snippet: Ectopic expression of A1CF inhibits EMT. ( a ) Overexpression A1CF and A1CF (-8AA) both result in gain of epithelial markers (E-cadherin) and loss of mesenchymal markers (vimentin and α-smooth muscle actin (α-SMA)), and the two variants have the similar trend in this process, GAPDH served as loading control; ( b ) Statistical analysis of relative protein expression levels. Data (mean ± standard error of the mean (S.E.M.)) are representative of three independent experiments. * p ≤ 0.05 and ** p ≤ 0.01 indicate significant statistical differences compared with the control group; ( c ) Immunofluorescence analysis demonstrates that ectopic expression of A1CF and A1CF (-8AA) potentiate the expression of membrane E-cadherin in NRK52e cells, and the expression of α-SMA and vimentin is weaker compared with control groups (original magnification, ×40). Scale bar represents 50 µm.

Article Snippet: SiRNA oligonucleotides were purchased from Shanghai GenePharma Co., Ltd. Two stealth siRNAs targeting different sequences of A1CF mRNA were selected to silence A1CF of Rattus norvegicus .

Techniques: Expressing, Over Expression, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Apobec-1 Complementation Factor (A1CF) Inhibits Epithelial-Mesenchymal Transition and Migration of Normal Rat Kidney Proximal Tubular Epithelial Cells

doi: 10.3390/ijms17020197

Figure Lengend Snippet: A1CF knockdown induces EMT. ( a ) Control-siRNA, A1CF-siRNA #1, and A1CF-siRNA #2 were transfected into NRK52e cells. Western blotting analysis reveals a decrease in E-cadherin (epithelial marker) and concomitant increase in vimentin and α-SMA (mesenchymal markers). GAPDH served as loading control; ( b ) Statistical analysis of relative protein expression levels , Data (mean ± S.E.M.) are representative of three independent experiments. ** p ≤ 0.01 indicates highly statistically significant differences compared with control-siRNA group; ( c ) A1CF-siRNA #2 was transfected into NRK52e cells. A1CF knockdown attenuates the expression of membrane E-cadherin and potentiates the expression of α-SMA and vimentin demonstrated by immunofluorescence analysis (original magnification, ×40). Scale bar represents 50 µm.

Article Snippet: SiRNA oligonucleotides were purchased from Shanghai GenePharma Co., Ltd. Two stealth siRNAs targeting different sequences of A1CF mRNA were selected to silence A1CF of Rattus norvegicus .

Techniques: Transfection, Western Blot, Marker, Expressing, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Apobec-1 Complementation Factor (A1CF) Inhibits Epithelial-Mesenchymal Transition and Migration of Normal Rat Kidney Proximal Tubular Epithelial Cells

doi: 10.3390/ijms17020197

Figure Lengend Snippet: A1CF inhibits NRK52e cells migration. ( a ) Both A1CF and A1CF (-8AA) significantly inhibited NRK52e cells migration. Data were collected at 12 and 24 h (original magnification, ×100); ( c ) A1CF knockdown significantly increases NRK52e cells migration. Data were collected at 12 and 24 h (original magnification, ×100); ( b , d ) Data are presented as mean ± S.E.M. of three independent experiments. Statistical analysis was performed using Student’s t -test. ** p < 0.01 indicated significantly statistical differences compared with control groups.

Article Snippet: SiRNA oligonucleotides were purchased from Shanghai GenePharma Co., Ltd. Two stealth siRNAs targeting different sequences of A1CF mRNA were selected to silence A1CF of Rattus norvegicus .

Techniques: Migration

Journal: Journal of Translational Medicine

Article Title: FNDC5 attenuates obesity-induced cardiac hypertrophy by inactivating JAK2/STAT3-associated inflammation and oxidative stress

doi: 10.1186/s12967-019-1857-8

Figure Lengend Snippet: FNDC5 deficiency aggravated HFD-induced inflammation in heart. a Cardiac Tnf - α, Il1b and Il6 mRNA levels determined with qPCR. b Phosphorylation levels of p38 and ERK in heart determined with Western blot. c NFκB activation in heart. N-p65: p65 in nucleus; C-p65: p65 in cytoplasm. Values are mean ± SEM. *P < 0.05 vs. WT. † P < 0.05 vs. Ctrl. n = 3

Article Snippet: The FNDC5 specific siRNA (sense sequence: 5′GAUGGCCUCUAAGAACAAA3′; antisense sequence: 3′CUACCGGAGAUUCUUGUUU5′) was purchased from RiboBio (Guangzhou, China).

Techniques: Western Blot, Activation Assay

Journal: Journal of Translational Medicine

Article Title: FNDC5 attenuates obesity-induced cardiac hypertrophy by inactivating JAK2/STAT3-associated inflammation and oxidative stress

doi: 10.1186/s12967-019-1857-8

Figure Lengend Snippet: FNDC5 deficiency enhanced HFD-induced oxidative stress and phosphorylated JAK2 and STAT3 level in heart. a Superoxide dismutase (SOD) activity and malondialdehyde (MDA) level in heart. b Expression of NAPDH oxidases (NOX2 and NOX4) protein in heart. c Phosphorylated JAK2 and STAT3 level in heart. Values are mean ± SEM. *P < 0.05 vs. WT. † P < 0.05 vs. Ctrl. n = 6

Article Snippet: The FNDC5 specific siRNA (sense sequence: 5′GAUGGCCUCUAAGAACAAA3′; antisense sequence: 3′CUACCGGAGAUUCUUGUUU5′) was purchased from RiboBio (Guangzhou, China).

Techniques: Activity Assay, Expressing

Journal: Journal of Translational Medicine

Article Title: FNDC5 attenuates obesity-induced cardiac hypertrophy by inactivating JAK2/STAT3-associated inflammation and oxidative stress

doi: 10.1186/s12967-019-1857-8

Figure Lengend Snippet: FNDC5 deficiency enhanced palmitate-induced inflammation and NOX4 expression in primary cardiomyocytes (CMs). a Effects of palmitate (PA, 400 μM) treatment on Tnf - α, Il1b and Il6 mRNA levels in CMs of WT and FNDC5 −/− mice. b Effects of PA treatment on NOX2 and NOX4 protein level in CMs of WT and FNDC5 −/− mice. The measurement was 24 h after PA treatment for measuring the mRNA levels or protein levels. PA: Palmitate; Veh: Vehicle; F −/− : FNDC5 −/− . Values are mean ± SEM. *P < 0.05 vs. Veh. † P < 0.05 vs. CMs-WT. n = 3

Article Snippet: The FNDC5 specific siRNA (sense sequence: 5′GAUGGCCUCUAAGAACAAA3′; antisense sequence: 3′CUACCGGAGAUUCUUGUUU5′) was purchased from RiboBio (Guangzhou, China).

Techniques: Expressing

Journal: Journal of Translational Medicine

Article Title: FNDC5 attenuates obesity-induced cardiac hypertrophy by inactivating JAK2/STAT3-associated inflammation and oxidative stress

doi: 10.1186/s12967-019-1857-8

Figure Lengend Snippet: Exogenous FNDC5 pretreatment attenuated palmitate-induced inflammation and oxidative stress in CMs. a , b Effects of exogenous FNDC5 pretreatment on cardiac hypertrophy markers ( Nppa, Nppb and Myh7 ) and inflammation markers ( Tnf - α, Il1b and Il6 ) mRNA levels in PA-stimulated CMs. c Effects of exogenous FNDC5 pretreatment on NO production in cell culture supernatant of PA-stimulated CMs. d Effects of exogenous FNDC5 pretreatment on NOX expression in PA-stimulated CMs. PA: Palmitate; Veh: Vehicle. Values are mean ± SEM. *P < 0.05 vs. Veh. † P < 0.05 vs. PA. n = 6

Article Snippet: The FNDC5 specific siRNA (sense sequence: 5′GAUGGCCUCUAAGAACAAA3′; antisense sequence: 3′CUACCGGAGAUUCUUGUUU5′) was purchased from RiboBio (Guangzhou, China).

Techniques: Cell Culture, Expressing

Journal: Journal of Translational Medicine

Article Title: FNDC5 attenuates obesity-induced cardiac hypertrophy by inactivating JAK2/STAT3-associated inflammation and oxidative stress

doi: 10.1186/s12967-019-1857-8

Figure Lengend Snippet: FNDC5 overexpression (OE) attenuated cardiac hypertrophy, inflammation and oxidative stress in HFD-fed mice. a , b Serum and heart/muscle FNDC5 levels after lentiviral vector-mediated FNDC5 overexpression. c Cardiomyocyte area in heart determined by H&E staining. d , e mRNA levels of the cardiac hypertrophy markers ( Nppa, Nppb and Myh7 ) and inflammation markers ( Tnf - α, Il1b and Il6 ). f SOD activity and MDA level in heart. g NOX2, NOX4 level and NFκB inactivation in heart. h Phosphorylation levels of p38 and ERK in heart. Values are mean ± SEM. *P < 0.05 vs. vector. n = 6

Article Snippet: The FNDC5 specific siRNA (sense sequence: 5′GAUGGCCUCUAAGAACAAA3′; antisense sequence: 3′CUACCGGAGAUUCUUGUUU5′) was purchased from RiboBio (Guangzhou, China).

Techniques: Over Expression, Plasmid Preparation, Staining, Activity Assay

Journal: Journal of Translational Medicine

Article Title: FNDC5 attenuates obesity-induced cardiac hypertrophy by inactivating JAK2/STAT3-associated inflammation and oxidative stress

doi: 10.1186/s12967-019-1857-8

Figure Lengend Snippet: Schematic illustrates a possible mechanism of FNDC5 in obesity-induced cardiac hypertrophy. Obesity-induced lipid overload has a great impact on the production of reactive oxygen species (ROS) and upregulation of TNF-α, IL-1β and IL-6 levels, which leads to cardiac inflammation and oxidative stress. The enhanced cardiac inflammation and oxidative stress are involved in pathogenesis of cardiac injury and remodeling. FNDC5 may exert its protective function on the enhanced inflammation and oxidative stress via JAK2/STAT3 pathway through an unknown receptor

Article Snippet: The FNDC5 specific siRNA (sense sequence: 5′GAUGGCCUCUAAGAACAAA3′; antisense sequence: 3′CUACCGGAGAUUCUUGUUU5′) was purchased from RiboBio (Guangzhou, China).

Techniques:

Journal: Journal of Translational Medicine

Article Title: FNDC5 attenuates obesity-induced cardiac hypertrophy by inactivating JAK2/STAT3-associated inflammation and oxidative stress

doi: 10.1186/s12967-019-1857-8

Figure Lengend Snippet: Echocardiographic assessment of left ventricle functions in mice

Article Snippet: The FNDC5 specific siRNA (sense sequence: 5′GAUGGCCUCUAAGAACAAA3′; antisense sequence: 3′CUACCGGAGAUUCUUGUUU5′) was purchased from RiboBio (Guangzhou, China).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: PFN1 is correlated with NSCLC metastasis and could promote NSCLC cell migration in vitro . (A) Representative IHC images of PFN1 expression on the NSCLC tissues. (B) The staining index of PFN1 on the tissue chip. ** p < 0.01. (C) Representative IHC images of PFN1 expression on the tissue chip. (D) The expression of PFN1 in TCGA LUAD data. ** p < 0.01. (E) The Kaplan–Meier survival analysis of PFN1 in NSCLC patients. (Data source: TCGA LUAD dataset) (F,G) Wound healing assays conducted to evaluate the migration ability of PFN1 -overexpressing (F) and PFN1 knockdown (KD) (G) H1299 cells. ** p < 0.01; scale bar, 500 μm. (H,I) Transwell migration assays conducted to evaluate the migration of PFN1 -overexpressing (H) and PFN1 KD (I) H1299 cells. ** p < 0.01; scale bar, 500 μm. EV, empty vector; OE, PFN1 overexpression; NC, negative control; si-1/ 2, PFN1 siRNA1 1/2.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: Migration, In Vitro, Expressing, Staining, Knockdown, Plasmid Preparation, Over Expression, Negative Control

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: PFN1 could promote MVs secretion in NSCLC. (A) Heatmap of differentially expressed proteins between EV and PFN1 OE cells. (B) GO enrichment analysis of differentially expressed proteins. (C) COG/KOG analysis of differentially expressed proteins. (D) MVs extracted from EV-expressing and PFN1 -overexpressing cells, using continuous differential centrifugation, identified using transmission electron microscopy. Scale bar, 100 nm. (E,F) Flow cytometry (E) and western blotting (F) were used to quantify MVs in PFN1 -overexpressing and EV-expressing cells. ARF6 and actin were used as MV markers. (G) Expression of PFN1 and annexin A1 in lung tumor tissues detected using immunofluorescence. (H) The staining index of p-MLC on the tissue chip. ** p < 0.01. (I) Representative IHC images of p-MLC expression. (J) Spearman rank correlation analysis was used to assess the relationship between PFN1 and p-MLC expression on the tissue chip; p and r values are shown in the plot.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: Expressing, Centrifugation, Transmission Assay, Electron Microscopy, Flow Cytometry, Western Blot, Immunofluorescence, Staining

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: MVs derived from PFN1 OE cells promote migration in NSCLC cells. (A) MVs collected from sera of patients with NSCLC quantified using flow cytometry. ** p < 0.01. (B) Protein expression of ARF6 and β-actin in MVs collected from sera of patients with NSCLC detected using western blotting. (C) Effect of PFN1 -overexpressing cell supernatants on cell migration evaluated through wound healing assays. ** p < 0.01; scale bar, 500 μm. (D) PKH67-labeled MVs taken up by H1299 cells. DAPI was used to stain the nuclei of H1299 cells. Scale bar, 500 μm. (E,F) Wound healing (E) and Transwell migration (F) assays conducted to evaluate the migration of H1299 cells after treatment with MVs derived from EV-expressing and PFN1 -overexpressing cells; ** p < 0.01; scale bar, 500 μm.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: Derivative Assay, Migration, Flow Cytometry, Expressing, Western Blot, Labeling, Staining

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: PFN1 promotes in vivo NSCLC metastasis by elevating MV secretion. (A) Schematic illustration of the mouse model of metastatic tumor established to determine the role of PFN1 in tumor metastasis. (B) Body weight changes in mice after intracardiac injection of PFN1 -overexpressing and EV-expressing cell lines. (C,D) Representative images of lung (C) and liver (D) metastases of the mouse model. The number of metastases is displayed in the right-hand side graph. * p < 0.05, ** p < 0.01. (E) Representative images of HE-stained lung tissues of the mouse model. (F) Representative IHC images of PFN1 and p-MLC expression in lung tissues. The staining index is shown in the right-hand side graph. ** p < 0.01. (G) Representative images of HE-stained liver tissues of the mouse model. (H) Representative IHC images of PFN1 and p-MLC expression in liver tissues. The staining index is shown in the right-hand side graph. ** p < 0.01. (I) Body weight changes in mice after intracardiac injection of H1299 cells and MVs. (J) Representative images of lung metastases of the mouse model. The number of metastases is shown in the bottom graph. * p < 0.05. (K) Representative images of HE-stained lung tissues of the mouse model. (L) Representative IHC images of PFN1 and p-MLC expression in lung tissues. The staining index is shown in the right-hand side graph; * p < 0.05.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: In Vivo, Injection, Expressing, Staining

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: Mechanisms underlying the promotion of MLC phosphorylation by PFN1. (A,B) Protein expression after PFN1 overexpression (A) and knockdown (B) measured using western blotting. (C) Protein expression in PFN1 mutants measured using western blotting. (D) PFN1 interactions with ROCK1/2 confirmed using co-IP. (E) Protein expression after treatment with Y27632 (10 µM) measured using western blotting. (F) Effect of PFN1 on ROCK1 activity. ** p < 0.01. (G) Effect of PFN1 on ROCK2 activity. (H) Flow cytometry measuring changes in the amount of MVs after treatment with Y27632; * p < 0.05.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: Phospho-proteomics, Expressing, Over Expression, Knockdown, Western Blot, Co-Immunoprecipitation Assay, Activity Assay, Flow Cytometry

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: ROCK1 inhibitor Y27632 partially reversed the promotion of lung cancer metastasis by PFN1 in vitro and in vivo . (A,B) Wound healing assays conducted to evaluate the effect of Y27632 (A) and Y27632 combined with MVs (B) on cell migration. ** p < 0.01; scale bar, 500 μm. (C) Transwell migration assays conducted to evaluate the effect of Y27632 and Y27632 combined with MVs on cell migration. ** p < 0.01; scale bar, 500 μm. (D) Schematic diagram of the mouse model of metastatic tumor established to determine the effect of Y27632 on PFN1-induced lung cancer metastasis. (E) Body weight changes in mice after intracardiac injection of PFN1 -overexpressing H1299 cells and intraperitoneal injection of Y27632 (10 mg/kg). (F) Representative images of lung and liver metastatic tissue in mice. The number of metastatic nodules is shown in the right-hand side graph. * p < 0.05. (G,H) Representative images of HE-stained lung (G) and liver (H) metastases. (I) Representative IHC images of PFN1 and p-MLC expression in lung tissues. The staining index is shown in the right-hand side graph. ** p < 0.01. (J) Representative IHC images of PFN1 and p-MLC expression in liver tissues. The staining index is shown in the right-hand side graph; ** p < 0.01.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: In Vitro, In Vivo, Migration, Injection, Staining, Expressing

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: Schematic diagram of the role of PFN1 in NSCLC metastasis. In the initiation stage of NSCLC, cells with upregulated PFN1 secret more MVs through PFN1 interactions with the ROCK/p-MLC pathway. These MVs contain numerous oncogenenic moleculars, which could enhance migration abilities of PFN1 normal expressed NSCLC cells, and untimately promote progression and metastasis of NSCLC.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: Migration