Journal: PLoS ONE

Article Title: Identifying Resistance Mechanisms against Five Tyrosine Kinase Inhibitors Targeting the ERBB/RAS Pathway in 45 Cancer Cell Lines

doi: 10.1371/journal.pone.0059503

Figure Lengend Snippet: Validation of the top genes by TaqMan RT-PCR in the cell lines.

Article Snippet: RAB17 , Hs00940833_m1 , 218931_at , RAB17, member RAS oncogene family , , 0.002.

Techniques: Biomarker Discovery, Binding Assay

Journal: Cell death & disease

Article Title: RAB17 promotes endometrial cancer progression by inhibiting TFRC-dependent ferroptosis.

doi: 10.1038/s41419-024-07013-w

Figure Lengend Snippet: Fig. 1 RAB17 regulates EC cell proliferation. A CCK-8 assays of Ishikawa and HEC-1A cell lines transfected with normal control siRNA (NC-si) or RAB17 siRNA (RAB17-si). B CCK-8 assays of Ishikawa and HEC-1A cells infected with normal control lentivirus (Lv-NC) or RAB17 overexpression lentivirus (Lv-RAB17). C Results from the EdU assays of Ishikawa and HEC-1A cells transfected with NC-si or RAB17-si for 48 h. Scale bars, 200 μm. D Results from the EdU assays of Ishikawa and HEC-1A cells infected with Lv-NC or Lv-RAB17 for 48 h. Scale bars, 200 μm. E Western blot analysis and (F) qRT–PCR analysis of RAB17 expression in Ishikawa and HEC-1A cells cultured in hyperglycemic (Hyper) or hypoglycemic (Hypo) medium for 72 h, respectively. GAPDH was used as an internal control. All the above assays were independently performed in triplicate (N = 3). The data are presented as the means ± SDs. The statistical analyses were performed by two-tailed unpaired Student’s t tests. **P < 0.01, and ***P < 0.001.

Article Snippet: The following primary antibodies were used: RAB17 (Proteintech, 17501-1-AP, 1:50 dilution) and TFRC (ABclonal, A5865, Wuhan, China, 1:100 dilution).

Techniques: CCK-8 Assay, Transfection, Control, Infection, Over Expression, Western Blot, Quantitative RT-PCR, Expressing, Cell Culture, Two Tailed Test

Journal: Cell death & disease

Article Title: RAB17 promotes endometrial cancer progression by inhibiting TFRC-dependent ferroptosis.

doi: 10.1038/s41419-024-07013-w

Figure Lengend Snippet: Fig. 3 RAB17 regulates TFRC expression in EC cells at the posttranscriptional level. A RAB17-binding proteins predicted based on the BioGRID, IntAct, MINT, ELM, and STRING databases. B Western blot analysis of TFRC expression in Ishikawa and HEC-1A cells transfected with NC-si or RAB17-si. C Western blot analysis of TFRC expression in Ishikawa and HEC-1A cells infected with Lv-NC or Lv-RAB17. D Representative images of IF staining showing TFRC expression in Ishikawa and HEC-1A cells transfected with NC-si or RAB17-si. Scale bars, 200 μm. E qRT–PCR analysis of TFRC expression in Ishikawa and HEC-1A cells transfected with NC-si or RAB17-si. F qRT–PCR analysis of TFRC expression in Ishikawa and HEC-1A cells infected with Lv-NC or Lv-RAB17. GAPDH was used as an internal control. All the above assays were independently performed in triplicate (N = 3). The data are presented as the means ± SDs. The statistical analyses were performed by two-tailed unpaired Student’s t tests. ***P < 0.001.

Article Snippet: The following primary antibodies were used: RAB17 (Proteintech, 17501-1-AP, 1:50 dilution) and TFRC (ABclonal, A5865, Wuhan, China, 1:100 dilution).

Techniques: Expressing, Binding Assay, Western Blot, Transfection, Infection, Staining, Quantitative RT-PCR, Control, Two Tailed Test

Journal: Cell death & disease

Article Title: RAB17 promotes endometrial cancer progression by inhibiting TFRC-dependent ferroptosis.

doi: 10.1038/s41419-024-07013-w

Figure Lengend Snippet: Fig. 5 RAB17 mediates TFRC-dependent ferroptosis in a hypoglycemic state. A Representative images of immunofluorescence staining with an ROS probe in Ishikawa cell lines cultured in hyperglycemic (Hyper) or hypoglycemic (Hypo) medium for the designated times. Scale bars, 200 μm. B GSH, SOD, MDA, and levels in Ishikawa and HEC-1A cells cultured in hyperglycemic (Hyper) or hypoglycemic (Hypo) medium for the designated times. C Western blot analysis of designated marker proteins for ferroptosis in Ishikawa and HEC-1A cells cultured in hyperglycemic (Hyper) or hypoglycemic (Hypo) medium for the designated times. D Western blot analysis of TFRC expression in Ishikawa cells cultured in hyperglycemic (Hyper) or hypoglycemic (Hypo) medium for the designated times. E Representative images of immunofluorescence staining for TFRC in Ishikawa cell lines cultured in hyperglycemic (Hyper) or hypoglycemic (Hypo) medium for the designated times. Scale bars, 200 μm. GAPDH was used as an internal control. All the above assays were independently performed in triplicate (N = 3). The data are presented as the means ± SDs. The statistical analyses were performed by two-tailed unpaired Student’s t tests. *P < 0.05, **P < 0.01, and ***P < 0.001.

Article Snippet: The following primary antibodies were used: RAB17 (Proteintech, 17501-1-AP, 1:50 dilution) and TFRC (ABclonal, A5865, Wuhan, China, 1:100 dilution).

Techniques: Staining, Cell Culture, Western Blot, Marker, Expressing, Control, Two Tailed Test

Journal: Cell death & disease

Article Title: RAB17 promotes endometrial cancer progression by inhibiting TFRC-dependent ferroptosis.

doi: 10.1038/s41419-024-07013-w

Figure Lengend Snippet: Fig. 6 TFRC-mediated ferroptosis is critical for RAB17-mediated regulation of EC cell proliferation. A Western blot analysis of RAB17 and TFRC expression in Ishikawa and HEC-1A cells cotransfected with the designated vectors. B CCK-8 assays of Ishikawa cell lines cotransfected with the designated vectors. C The iron contents of Ishikawa cell lines cotransfected with the designated vectors. D Representative images of immunofluorescence staining with an ROS probe in Ishikawa cell lines cotransfected with the designated vectors. Scale bars, 200 μm. E Western blot analysis of RAB17 and TFRC expression in Ishikawa and HEC-1A cells transfected with/without designated siRNAs or treated with/without Fer-1. F CCK-8 assays of Ishikawa cell lines transfected with/without designated siRNAs or treated with/without Fer-1. G Iron content of Ishikawa cell lines transfected with/without designated siRNAs or treated with/without Fer-1. H Representative images of immunofluorescence staining using an ROS probe in Ishikawa cell lines transfected with/without the indicated siRNAs or treated with/without Fer-1. Scale bars, 200 μm. GAPDH was used as an internal control. All the above assays were independently performed in triplicate (N = 3). The data are presented as the means ± SDs. The statistical analyses were performed by two-tailed unpaired Student’s t tests. **P < 0.01, and ***P < 0.001.

Article Snippet: The following primary antibodies were used: RAB17 (Proteintech, 17501-1-AP, 1:50 dilution) and TFRC (ABclonal, A5865, Wuhan, China, 1:100 dilution).

Techniques: Western Blot, Expressing, CCK-8 Assay, Staining, Transfection, Control, Two Tailed Test

Journal: Cell death & disease

Article Title: RAB17 promotes endometrial cancer progression by inhibiting TFRC-dependent ferroptosis.

doi: 10.1038/s41419-024-07013-w

Figure Lengend Snippet: Fig. 7 RAB17 regulates ferroptosis in ECs in vivo. A Representative images of the xenografts, (B) tumor volume, and (C) tumor weight 31 days after inoculation of Ishikawa cells infected with Lv-NC or Lv-RAB17 (n = 6 per group). The tumor volumes were measured every 3 days. D Western blot analysis of RAB17 and TFRC expression in Lv-NC and Lv-RAB17 xenografts. E Representative images of immunohistochemical staining for RAB17, TFRC, and KI67, as well as Prussian blue staining, of Lv-NC and Lv-RAB17 xenografts. Scale bars, 100 μm. F ROS levels in Lv-NC and Lv-RAB17 xenografts. G GSH, SOD, MDA, and iron levels in Lv-NC and Lv-RAB17 xenografts. H Western blot analysis of designated ferroptosis marker proteins in Lv-NC and Lv-RAB17 xenografts. GAPDH was used as an internal control. H Representative images of the xenografts, (I) tumor volume, and weight 28 days after inoculation of Ishikawa cells infected with siNC or siRAB17 (n = 5 per group). The tumor volumes were measured every 3 days. J Representative images of immunohistochemical staining for RAB17, TFRC, and KI67, as well as Prussian blue staining, of siNC or siRAB17 xenografts. Scale bars, 100 μm. K Representative images of the xenografts, tumor volume, and weight 28 days after inoculation of Ishikawa cells infected with indicated vectors (n = 5 per group). The tumor volumes were measured every 3 days. L Representative images of immunohistochemical staining for RAB17, TFRC, and KI67, as well as Prussian blue staining, of indicated xenografts. Scale bars, 100 μm. All the above assays were independently performed in triplicate (N = 3). The data are presented as the means ± SDs. The statistical analyses were performed by two-tailed unpaired Student’s t tests. **P < 0.01, and ***P < 0.001.

Article Snippet: The following primary antibodies were used: RAB17 (Proteintech, 17501-1-AP, 1:50 dilution) and TFRC (ABclonal, A5865, Wuhan, China, 1:100 dilution).

Techniques: In Vivo, Infection, Western Blot, Expressing, Immunohistochemical staining, Staining, Marker, Control, Two Tailed Test

Journal: Cell death & disease

Article Title: RAB17 promotes endometrial cancer progression by inhibiting TFRC-dependent ferroptosis.

doi: 10.1038/s41419-024-07013-w

Figure Lengend Snippet: Fig. 8 RAB17 and TFRC expression levels are significantly associated with prognosis in patients with EC. A Representative IHC images of RAB17 and TFRC expression in 118 EC tissues. The right panels (scale bars = 25 μm) show magnified views of the boxed area in the corresponding left panels (scale bars = 300 μm). B Chi-square test based on the immunohistochemical analysis of RAB17 and TFRC expression. C The overall survival (OS) of EC patients with different RAB17 protein expression levels were assessed by Kaplan–Meier survival curves and log-rank tests. D Multivariate analysis of factors associated with overall survival in patients with EC. E Representative immunofluorescence images of RAB17 and TFRC expression in 86 EC tissues from our cohort. F Correlation analysis between RAB17 and TFRC expression, chi-square test was used. G The overall survival (OS) of 86 EC patients from our cohort with different RAB17 protein expression levels were assessed by Kaplan–Meier survival curves and log-rank tests. ***P < 0.001.

Article Snippet: The following primary antibodies were used: RAB17 (Proteintech, 17501-1-AP, 1:50 dilution) and TFRC (ABclonal, A5865, Wuhan, China, 1:100 dilution).

Techniques: Expressing, Immunohistochemical staining

Journal: European journal of pharmacology

Article Title: Pterostilbene inhibits melanogenesis, melanocyte dendricity and melanosome transport through cAMP/PKA/CREB pathway.

doi: 10.1016/j.ejphar.2022.175231

Figure Lengend Snippet: Fig. 5. Pterostilbene inhibits dendritic formation and melanosome transport in B16F10 cells. (A) Treated with pterostilbene (0.1, 1, and 3 μM) for 72 h in B16F10 cells, Stained with FITC-Phalloidin fluorescence probe to visualize the cytoskeleton (green) and the Cell nuclei were counterstained with DAPI (blue). Cytoskeleton changes in B16F10 cells were observed by fluorescence microscope at 400 × magnification, Bar = 50 μm. (B–F) Treated with pterostilbene (0.1, 1, and 3 μM) for 12 h in B16F10 cells, and detected Rab27A (B), MLPH (C), Myosin Va (D), Rab17 (E) and gp100 (F) gene expression by RT-qPCR. (G) Treated with pterostilbene (0.1, 1, and 3 μM) for 72 h in B16F10 cells, and WB was then applied to detect the protein levels of Rab27A, MLPH, Myosin Va, Rab17 and gp100. Data are expressed as the mean ± SEM (n = 3). *p < 0.05, **p < 0.01 versus non-treated cells.

Article Snippet: Antibodies against β-actin (3700S, 1:1000), Rab27A (69295S, 1:1000), Myosin Va (3402S, 1:1000), Rab17 (8013S, 1:1000), Second antibody anti-mouse IgG (7076S, 1:1000) and anti-rabbit IgG (7074S, 1:1000) were obtained from Cell Signaling Technology (USA).

Techniques: Staining, Fluorescence, Microscopy, Gene Expression, Quantitative RT-PCR

Journal: European journal of pharmacology

Article Title: Pterostilbene inhibits melanogenesis, melanocyte dendricity and melanosome transport through cAMP/PKA/CREB pathway.

doi: 10.1016/j.ejphar.2022.175231

Figure Lengend Snippet: Fig. 7. Effects of HY-B0764 on pterostilbene-inhibited melanogenesis and melanosome transport of B16F10 cells. B16F10 cells was pretreated or not with 3 μM HY- B0764 for 1 h before pterostilbene was applied for 24 h, 48h or 72 h at 3 μM. (A) The CREB and p-CREB protein expression levels were detected by WB (B–D) B16F10 cells were stained with Masson-Fontana ammoniacal silver stain. Bar = 40 μm (B), Number of dendrites per cell was counted (C); Length of dendrites per cell was measured(D). (E) WB for the protein expression relating to melanogenesis (MITF, TYR) and melanosome transport (Rab27A, Rab17, gp100) were detected. Results shown are the mean ± SEM (n = 9). *p < 0.05, **p < 0.01 versus non-treated cells. #p < 0.05 versus pterostilbene-treated cells.

Article Snippet: Antibodies against β-actin (3700S, 1:1000), Rab27A (69295S, 1:1000), Myosin Va (3402S, 1:1000), Rab17 (8013S, 1:1000), Second antibody anti-mouse IgG (7076S, 1:1000) and anti-rabbit IgG (7074S, 1:1000) were obtained from Cell Signaling Technology (USA).

Techniques: Expressing, Staining, Silver Staining

Journal: OncoTargets and therapy

Article Title: Knockdown of Circular RNA Hsa_circ_0000714 Can Regulate RAB17 by Sponging miR-370-3p to Reduce Paclitaxel Resistance of Ovarian Cancer Through CDK6/RB Pathway

doi: 10.2147/OTT.S285153

Figure Lengend Snippet: Primer Sequences Used for siRNA

Article Snippet: Lentiviruses carrying RAB17 expression vectors were obtained from GeneChem (Shanghai, China).

Techniques:

Journal: OncoTargets and therapy

Article Title: Knockdown of Circular RNA Hsa_circ_0000714 Can Regulate RAB17 by Sponging miR-370-3p to Reduce Paclitaxel Resistance of Ovarian Cancer Through CDK6/RB Pathway

doi: 10.2147/OTT.S285153

Figure Lengend Snippet: Primer Sequences Used for qRT-PCR

Article Snippet: Lentiviruses carrying RAB17 expression vectors were obtained from GeneChem (Shanghai, China).

Techniques: Sequencing

Journal: OncoTargets and therapy

Article Title: Knockdown of Circular RNA Hsa_circ_0000714 Can Regulate RAB17 by Sponging miR-370-3p to Reduce Paclitaxel Resistance of Ovarian Cancer Through CDK6/RB Pathway

doi: 10.2147/OTT.S285153

Figure Lengend Snippet: Higher RAB17 expression in paclitaxel-resistant ovarian cancer cells than in paclitaxel-sensitive ovarian cancer cells. ( A ) The IC 50 of paclitaxel-sensitive ovarian cancer cells SKOV3, A2780, and paclitaxel-resistant ovarian cancer cells SKOV3/PTX, A2780/PTX. ( B ) Expression of RAB17 in paclitaxel-sensitive ovarian cancer cells SKOV3, A2780, and paclitaxel-resistant ovarian cancer cells SKOV3/PTX, A2780/PTX, *p<0.05.

Article Snippet: Lentiviruses carrying RAB17 expression vectors were obtained from GeneChem (Shanghai, China).

Techniques: Expressing

Journal: OncoTargets and therapy

Article Title: Knockdown of Circular RNA Hsa_circ_0000714 Can Regulate RAB17 by Sponging miR-370-3p to Reduce Paclitaxel Resistance of Ovarian Cancer Through CDK6/RB Pathway

doi: 10.2147/OTT.S285153

Figure Lengend Snippet: The Up-Regulated and Down-Regulated Genes of A2780/PTX vs A2780 are Arranged According to the Ratio Change of the Microarray Data

Article Snippet: Lentiviruses carrying RAB17 expression vectors were obtained from GeneChem (Shanghai, China).

Techniques: Microarray

Journal: OncoTargets and therapy

Article Title: Knockdown of Circular RNA Hsa_circ_0000714 Can Regulate RAB17 by Sponging miR-370-3p to Reduce Paclitaxel Resistance of Ovarian Cancer Through CDK6/RB Pathway

doi: 10.2147/OTT.S285153

Figure Lengend Snippet: Knockdown and overexpression of RAB17 in A2780/PTX and A2780 cells, respectively, influence paclitaxel resistance, proliferation, cell cycle, and protein changes. ( A ) The transfection efficiency of knockdown and overexpression of RAB17 in A2780/PTX and A2780 cells, respectively, were identified by qRT-PCR and Western blotting, **p<0.01. ( B ) Influence of paclitaxel resistance following knockdown and overexpression of RAB17 in A2780/PTX and A2780 cells, respectively, as detected by CCK-8. ( C ) Influence of cell proliferation following knockdown and overexpression of RAB17 in A2780/PTX and A2780 cells, respectively, as detected by colony formation assays, **p<0.01. ( D ) Influence of the cell cycle following knockdown and overexpression of RAB17 in A2780/PTX and A2780 cells, respectively, as detected by flow cytometry assay, *p<0.05, **p<0.01. ( E ) Influence of apoptotic and adhesion related proteins following knockdown and overexpression RAB17 in A2780/PTX and A2780 cells, respectively, as detected by Western blotting, *p<0.05, **p<0.01.

Article Snippet: Lentiviruses carrying RAB17 expression vectors were obtained from GeneChem (Shanghai, China).

Techniques: Knockdown, Over Expression, Transfection, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Flow Cytometry

Journal: OncoTargets and therapy

Article Title: Knockdown of Circular RNA Hsa_circ_0000714 Can Regulate RAB17 by Sponging miR-370-3p to Reduce Paclitaxel Resistance of Ovarian Cancer Through CDK6/RB Pathway

doi: 10.2147/OTT.S285153

Figure Lengend Snippet: RAB17 regulates a variety of cellular behaviors in A2780/PTX and A2780 cells by activating the CDK6/RB signaling pathway. ( A ) Expression of P-gp protein was observed after knockdown and overexpression of RAB17 in A2780/PTX and A2780 cells respectively, *p<0.05, **p<0.01. ( B ) Expression of cell cycle related proteins was observed after knockdown and overexpression RAB17 in A2780/PTX and A2780 cells respectively, **p<0.01.

Article Snippet: Lentiviruses carrying RAB17 expression vectors were obtained from GeneChem (Shanghai, China).

Techniques: Expressing, Knockdown, Over Expression

Journal: OncoTargets and therapy

Article Title: Knockdown of Circular RNA Hsa_circ_0000714 Can Regulate RAB17 by Sponging miR-370-3p to Reduce Paclitaxel Resistance of Ovarian Cancer Through CDK6/RB Pathway

doi: 10.2147/OTT.S285153

Figure Lengend Snippet: RAB17 is a new target of miR-370-3p. ( A ) The putative binding sites of miR-370-3p and RAB17. ( B ) Luciferase activities of wild-type (WT) RAB17 3′-UTR and mutant-type (MUT) RAB17 3ʹ-UTR after transfection with miR-370-3p mimics, **p<0.01.

Article Snippet: Lentiviruses carrying RAB17 expression vectors were obtained from GeneChem (Shanghai, China).

Techniques: Binding Assay, Luciferase, Mutagenesis, Transfection

Journal: OncoTargets and therapy

Article Title: Knockdown of Circular RNA Hsa_circ_0000714 Can Regulate RAB17 by Sponging miR-370-3p to Reduce Paclitaxel Resistance of Ovarian Cancer Through CDK6/RB Pathway

doi: 10.2147/OTT.S285153

Figure Lengend Snippet: Influence of overexpressing miR-370-3p on paclitaxel resistance, cell proliferation, cell cycle, and signaling pathway in A2780/PTX. ( A ) The expression level of RAB17 after A2780/PTX cell transfection with miR-370-3p mimics, **p<0.01. ( B ) Influence of miR-370-3p transfection with mimics on paclitaxel resistance in A2780/PTX cell detected by CCK-8. ( C ) Influence of miR-370-3p transfection with mimics on cell proliferation in A2780/PTX cells assessed by colony formation assays, **p<0.01. ( D ) Influence of miR-370-3p transfection with mimics on the cell cycle in A2780/PTX cell detected by flow cytometry assay, *p<0.05, **p<0.01. ( E ) Influence of miR-370-3p transfection with mimics on the CDK6/RB signaling pathway proteins in A2780/PTX cells detected by Western blotting, **p<0.01.

Article Snippet: Lentiviruses carrying RAB17 expression vectors were obtained from GeneChem (Shanghai, China).

Techniques: Expressing, Transfection, CCK-8 Assay, Flow Cytometry, Western Blot

Journal: OncoTargets and therapy

Article Title: Knockdown of Circular RNA Hsa_circ_0000714 Can Regulate RAB17 by Sponging miR-370-3p to Reduce Paclitaxel Resistance of Ovarian Cancer Through CDK6/RB Pathway

doi: 10.2147/OTT.S285153

Figure Lengend Snippet: Hsa_circ_0000714 acts as a molecular sponge of miR-370-3p. ( A ) Hsa_circ_0000714 expression was remarkably higher in A2780/PTX cells compared with other circRNAs, **p<0.01. ( B ) Transfection with specific siRNAs, including si-hsa_circ_0000714#1 and si-hsa_circ_0000714#2, showing knockdown of hsa_circ_0000714 expression, **p<0.01. ( C ) Knockdown of hsa_circ_0000714 upregulated miR-370-3p expression and further downregulated RAB17 expression, **p<0.01. ( D ) Putative binding sites between hsa_circ_0000714 and miR-370-3p. ( E ) Overexpressed miR-370-3p reduced the luciferase activity of wild-type (WT) hsa_circ_0000714 compared with mutant-type (MUT) hsa_circ_0000714 as detected by luciferase reporter assay, *p<0.05.

Article Snippet: Lentiviruses carrying RAB17 expression vectors were obtained from GeneChem (Shanghai, China).

Techniques: Expressing, Transfection, Knockdown, Binding Assay, Luciferase, Activity Assay, Mutagenesis, Reporter Assay

Journal: OncoTargets and therapy

Article Title: Knockdown of Circular RNA Hsa_circ_0000714 Can Regulate RAB17 by Sponging miR-370-3p to Reduce Paclitaxel Resistance of Ovarian Cancer Through CDK6/RB Pathway

doi: 10.2147/OTT.S285153

Figure Lengend Snippet: Schematic model illustrating the biological roles of the hsa_circ_0000714/miR-370-3p/RAB17 regulatory axis in A2780/PTX paclitaxel-resistant ovarian cancer cell.

Article Snippet: Lentiviruses carrying RAB17 expression vectors were obtained from GeneChem (Shanghai, China).

Techniques:

Journal: Burns & Trauma

Article Title: Single-cell RNA-seq and bulk-seq identify RAB17 as a potential regulator of angiogenesis by human dermal microvascular endothelial cells in diabetic foot ulcers

doi: 10.1093/burnst/tkad020

Figure Lengend Snippet: RAB17 level was decreased under hyperglycemic conditions. RAB17 expression level in HDMECs between normal and DFU group was measured by ( a ) q-PCR and ( b ) western blotting. ( c , d ) Western blotting assessing the expression of RAB17 in Nor-HDMEC exposed to high-glucose medium (30 mM D-glucose) for 2 and 7 days; 30 mM mannitol was used as an osmotic control. ( e ) Immunohistochemical analysis of RAB17 in the skin of normal patients and DFU subjects. Scale bar: 200 μm (upper panel) and 40 μm (lower panel) respectively. ( f ) Immunohistochemical analysis of RAB17 in the skin of normal mice and diabetic mice. Scale bar: 200 μm (upper panel) and 40 μm (lower panel) respectively. Results are represented as the mean ± SD; * p < 0.05, * * p < 0.01. DFU diabetic foot ulcer, Nor normal, HDMECs human dermal microvascular endothelial cells, HG high glucose, SD standard deviation

Article Snippet: HDMECs were seeded in 6-well plates and cultured to 80% confluence. siRNA targeting RAB17 (si-RAB17-1, si-RAB17-2) and scrambled negative control (si-NC) were procured from RiboBio (Guangzhou, Guangdong, China).

Techniques: Expressing, Western Blot, Control, Immunohistochemical staining, Standard Deviation

Journal: Burns & Trauma

Article Title: Single-cell RNA-seq and bulk-seq identify RAB17 as a potential regulator of angiogenesis by human dermal microvascular endothelial cells in diabetic foot ulcers

doi: 10.1093/burnst/tkad020

Figure Lengend Snippet: RAB17 affected angiogenic capacity of HDMECs in vitro . ( a , b ) Western blotting assessing the expression of RAB17 in DFU-HDMECs after RAB17-overexpressing lentivirus transfection, and the expressions of HIF-1α, VEGF-A, ERK and p-ERK in DFU-HDMECs after overexpression of RAB17. ( c ) Matrigel tube-formation assay of DFU-HDMECs following RAB17 overexpression and the quantitative analysis of tube formation using length of tubes and number of nodes. ( d , e ) Western blotting assessing the expressions of RAB17, HIF-1α, VEGF-A, ERK and p-ERK after knockdown by siRNA targeting RAB17. ( f ) Matrigel tube-formation assay and quantitative analysis of Nor-HDMECs following knock-down of RAB17. ( g , h ) Western blotting revealing the expression levels of HIF-1α, VEGF-A, ERK and p-ERK after overexpression of RAB17 with or without PD98059. ( i ) Matrigel tube-formation assay and quantitative analysis of DFU-HDMECs following RAB17 overexpression with or without PD98059. Results are represented as the mean ± SD; * p < 0.05, * * p < 0.01. Lv lentivirus, Ctrl control, DFU diabetic foot ulcer, Nor normal, HDMECs human dermal microvascular endothelial cells, HIF-1α hypoxia inducible factor 1 subunit alpha, VEGF-A vascular endothelial growth factor A, si-RAB17 RAB17 small interfering RNA, si-NC negative control small interfering RNA, SD standard deviation

Article Snippet: HDMECs were seeded in 6-well plates and cultured to 80% confluence. siRNA targeting RAB17 (si-RAB17-1, si-RAB17-2) and scrambled negative control (si-NC) were procured from RiboBio (Guangzhou, Guangdong, China).

Techniques: In Vitro, Western Blot, Expressing, Transfection, Over Expression, Tube Formation Assay, Knockdown, Control, Small Interfering RNA, Negative Control, Standard Deviation

Journal: Burns & Trauma

Article Title: Single-cell RNA-seq and bulk-seq identify RAB17 as a potential regulator of angiogenesis by human dermal microvascular endothelial cells in diabetic foot ulcers

doi: 10.1093/burnst/tkad020

Figure Lengend Snippet: RAB17 overexpression promoted diabetic angiogenesis and wound healing in vivo . ( a ) Immunohistochemical analysis of RAB17 between groups after injection of rAAV-RAB17 or rAAV-vector. Scale bar: 40 μm. ( b , c ) Western blotting assessing the expression of RAB17 in rAAV-RAB17 or rAAV-vector transfected mice skin. ( d ) Laser speckle contrast imager recording the wound perfusion images and ( e ) relative wound perfusion ratio in rAAV-RAB17 group and the control vector group; high- or low blood flow are represented in red and blue respectively. ( f ) Wound area monitored at the indicated days post-wounding and ( g ) quantitative analysis in rAAV-RAB17 group and the control vector group. Results are represented as the mean ± SD; * p < 0.05. rAAV recombinant adeno-associated viral, SD standard deviation

Article Snippet: HDMECs were seeded in 6-well plates and cultured to 80% confluence. siRNA targeting RAB17 (si-RAB17-1, si-RAB17-2) and scrambled negative control (si-NC) were procured from RiboBio (Guangzhou, Guangdong, China).

Techniques: Over Expression, In Vivo, Immunohistochemical staining, Injection, Plasmid Preparation, Western Blot, Expressing, Transfection, Control, Recombinant, Standard Deviation

Journal: Burns & Trauma

Article Title: Single-cell RNA-seq and bulk-seq identify RAB17 as a potential regulator of angiogenesis by human dermal microvascular endothelial cells in diabetic foot ulcers

doi: 10.1093/burnst/tkad020

Figure Lengend Snippet: RAB17 level was decreased under hyperglycemic conditions. RAB17 expression level in HDMECs between normal and DFU group was measured by ( a ) q-PCR and ( b ) western blotting. ( c , d ) Western blotting assessing the expression of RAB17 in Nor-HDMEC exposed to high-glucose medium (30 mM D-glucose) for 2 and 7 days; 30 mM mannitol was used as an osmotic control. ( e ) Immunohistochemical analysis of RAB17 in the skin of normal patients and DFU subjects. Scale bar: 200 μm (upper panel) and 40 μm (lower panel) respectively. ( f ) Immunohistochemical analysis of RAB17 in the skin of normal mice and diabetic mice. Scale bar: 200 μm (upper panel) and 40 μm (lower panel) respectively. Results are represented as the mean ± SD; * p < 0.05, * * p < 0.01. DFU diabetic foot ulcer, Nor normal, HDMECs human dermal microvascular endothelial cells, HG high glucose, SD standard deviation

Article Snippet: Immunohistochemistry staining was performed using an anti-RAB17 antibody (1 : 200, Zenbio, Chengdu, China) and incubated overnight at 4°C.

Techniques: Expressing, Western Blot, Control, Immunohistochemical staining, Standard Deviation

Journal: Burns & Trauma

Article Title: Single-cell RNA-seq and bulk-seq identify RAB17 as a potential regulator of angiogenesis by human dermal microvascular endothelial cells in diabetic foot ulcers

doi: 10.1093/burnst/tkad020

Figure Lengend Snippet: RAB17 affected angiogenic capacity of HDMECs in vitro . ( a , b ) Western blotting assessing the expression of RAB17 in DFU-HDMECs after RAB17-overexpressing lentivirus transfection, and the expressions of HIF-1α, VEGF-A, ERK and p-ERK in DFU-HDMECs after overexpression of RAB17. ( c ) Matrigel tube-formation assay of DFU-HDMECs following RAB17 overexpression and the quantitative analysis of tube formation using length of tubes and number of nodes. ( d , e ) Western blotting assessing the expressions of RAB17, HIF-1α, VEGF-A, ERK and p-ERK after knockdown by siRNA targeting RAB17. ( f ) Matrigel tube-formation assay and quantitative analysis of Nor-HDMECs following knock-down of RAB17. ( g , h ) Western blotting revealing the expression levels of HIF-1α, VEGF-A, ERK and p-ERK after overexpression of RAB17 with or without PD98059. ( i ) Matrigel tube-formation assay and quantitative analysis of DFU-HDMECs following RAB17 overexpression with or without PD98059. Results are represented as the mean ± SD; * p < 0.05, * * p < 0.01. Lv lentivirus, Ctrl control, DFU diabetic foot ulcer, Nor normal, HDMECs human dermal microvascular endothelial cells, HIF-1α hypoxia inducible factor 1 subunit alpha, VEGF-A vascular endothelial growth factor A, si-RAB17 RAB17 small interfering RNA, si-NC negative control small interfering RNA, SD standard deviation

Article Snippet: Immunohistochemistry staining was performed using an anti-RAB17 antibody (1 : 200, Zenbio, Chengdu, China) and incubated overnight at 4°C.

Techniques: In Vitro, Western Blot, Expressing, Transfection, Over Expression, Tube Formation Assay, Knockdown, Control, Small Interfering RNA, Negative Control, Standard Deviation

Journal: Burns & Trauma

Article Title: Single-cell RNA-seq and bulk-seq identify RAB17 as a potential regulator of angiogenesis by human dermal microvascular endothelial cells in diabetic foot ulcers

doi: 10.1093/burnst/tkad020

Figure Lengend Snippet: RAB17 overexpression promoted diabetic angiogenesis and wound healing in vivo . ( a ) Immunohistochemical analysis of RAB17 between groups after injection of rAAV-RAB17 or rAAV-vector. Scale bar: 40 μm. ( b , c ) Western blotting assessing the expression of RAB17 in rAAV-RAB17 or rAAV-vector transfected mice skin. ( d ) Laser speckle contrast imager recording the wound perfusion images and ( e ) relative wound perfusion ratio in rAAV-RAB17 group and the control vector group; high- or low blood flow are represented in red and blue respectively. ( f ) Wound area monitored at the indicated days post-wounding and ( g ) quantitative analysis in rAAV-RAB17 group and the control vector group. Results are represented as the mean ± SD; * p < 0.05. rAAV recombinant adeno-associated viral, SD standard deviation

Article Snippet: Immunohistochemistry staining was performed using an anti-RAB17 antibody (1 : 200, Zenbio, Chengdu, China) and incubated overnight at 4°C.

Techniques: Over Expression, In Vivo, Immunohistochemical staining, Injection, Plasmid Preparation, Western Blot, Expressing, Transfection, Control, Recombinant, Standard Deviation