Journal: Journal of Cellular and Molecular Medicine

Article Title: Macrophage‐derived exosomes mediate silica‐induced pulmonary fibrosis by activating fibroblast in an endoplasmic reticulum stress‐dependent manner

doi: 10.1111/jcmm.16524

Figure Lengend Snippet: Macrophage‐derived exosomes modulate myofibroblast differentiation in vitro. (A) Exosomes derived from RAW264.7 cells were labelled with PKH26 dye and then incubated with HFL1 cells for 24 h. Scale bar: 20 μm. (B, D‐E) Western blot (B) and RT‐qPCR (D‐E) analyses of collagen I, α‐SMA and GAPDH in HFL1 cells treated with PBS, NC‐Exos, SiO 2 ‐Exos or SiO 2 + GW4869‐Exos were performed. These exosomes were derived from THP‐1 cells. (C, F‐G) Western blot (C) and RT‐qPCR (F‐G) analyses of collagen I, α‐SMA and GAPDH in NIH‐3T3 cells incubated with PBS, NC‐Exos, SiO 2 ‐Exos or SiO 2 + GW4869‐Exos were performed. These exosomes were derived from RAW264.7 cells. Student's t test; * P < .05, ** P < .01

Article Snippet: The mouse macrophage cell line RAW264.7, mouse embryonic fibroblast cell line NIH‐3T3, human monocyte leukaemia cell line THP‐1 and human embryonic fibroblast cell line HFL1 were purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Derivative Assay, In Vitro, Incubation, Western Blot, Quantitative RT-PCR

Journal: Journal of Cellular and Molecular Medicine

Article Title: Macrophage‐derived exosomes mediate silica‐induced pulmonary fibrosis by activating fibroblast in an endoplasmic reticulum stress‐dependent manner

doi: 10.1111/jcmm.16524

Figure Lengend Snippet: SiO 2 ‐Exos induce lung fibroblast proliferation and migration (A‐B) CCK‐8 assay was used to evaluate the viability of HFL1 cells (A) and NIH‐3T3 cells (B) treated with various exosomes (SiO 2 ‐Exos or SiO 2 + GW4869‐Exos). Two‐way ANOVA; ** P < .01, *** P < .001, n.s: not significant. (C‐D) The migration of HFL1 cells (C) and NIH‐3T3 cells (D) was assessed by wound closure assay. Scale bar: 100 μm. Student's t test; * P < .05, ** P < .01, *** P < .001, n.s: not significant

Article Snippet: The mouse macrophage cell line RAW264.7, mouse embryonic fibroblast cell line NIH‐3T3, human monocyte leukaemia cell line THP‐1 and human embryonic fibroblast cell line HFL1 were purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Migration, CCK-8 Assay, Wound Closure Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Macrophage‐derived exosomes mediate silica‐induced pulmonary fibrosis by activating fibroblast in an endoplasmic reticulum stress‐dependent manner

doi: 10.1111/jcmm.16524

Figure Lengend Snippet: SiO 2 ‐Exo‐induced myofibroblast differentiation is ER stress dependent (A‐C, E) Western blot analysis of BIP, XBP1s, P ‐eIF2α, α‐SMA, Collagen I, and GAPDH in HFL1 cells (A, C) and NIH‐3T3 cells (B, E) incubated with SiO 2 ‐Exos (± 4‐PBA) at different points in time (0, 8, 16 or 24 h). (D, F) Analysis of collagen I, α‐SMA, and GAPDH in HFL1 cells (D) and NIH‐3T3 cells (F) incubated with SiO 2 ‐Exos or SiO 2 ‐Exos +4‐PBA for 24 h

Article Snippet: The mouse macrophage cell line RAW264.7, mouse embryonic fibroblast cell line NIH‐3T3, human monocyte leukaemia cell line THP‐1 and human embryonic fibroblast cell line HFL1 were purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Western Blot, Incubation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Macrophage‐derived exosomes mediate silica‐induced pulmonary fibrosis by activating fibroblast in an endoplasmic reticulum stress‐dependent manner

doi: 10.1111/jcmm.16524

Figure Lengend Snippet: Inhibition of ER stress attenuates SiO 2 ‐Exo‐induced lung fibroblast proliferation and migration (A‐B) CCK‐8 assay was used to evaluate the viability of HFL1 cells (A) and NIH‐3T3 cells (B) incubated with SiO 2 ‐Exos ± 4‐PBA. Two‐way ANOVA; * P < .05, ** P < .01, *** P < .001, n.s: not significant. (C‐D) The migration of HFL1 cells (C) and NIH‐3T3 cells (D) was assessed by wound closure assay. Scale bar: 100 μm. Student's t test; * P < .05, ** P < .01, *** P < .001, n.s: not significant

Article Snippet: The mouse macrophage cell line RAW264.7, mouse embryonic fibroblast cell line NIH‐3T3, human monocyte leukaemia cell line THP‐1 and human embryonic fibroblast cell line HFL1 were purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Inhibition, Migration, CCK-8 Assay, Incubation, Wound Closure Assay

Journal: Molecular Oncology

Article Title: Cancer‐associated fibroblast migration in non‐small cell lung cancers is modulated by increased integrin α11 expression

doi: 10.1002/1878-0261.12937

Figure Lengend Snippet: Effect of ITGA11 genetic modification on fibroblast features. (A) ITGA11 was knocked down using siRNA in HFL‐1 and proteins were extracted and subjected to western blot analysis to detect the expression levels of each target (see Section ). Vertical axis: (B) ITGA11, (C) COL11A1, (D) integrin α5, (E) integrin β1, (F) fibronectin, and (G) α‐SMA protein expression normalized to β‐actin, (H) p‐ERK1/2 protein expression normalized to t‐ERK1/2 and (I) p‐SMAD3 protein expression normalized to t‐SMAD3. (J) Migration toward fibronectin (20 µg·mL −1 ) was assessed after silencing ITGA11 . Vertical axis: number of migrated cells per 5HPF. (K) Migration toward collagen type I (1 µg·mL −1 ) was assessed after silencing ITGA11 . (B‐K; n ≥ 3, one‐way ANOVA) Vertical axis: number of migrated cells per 5HPF. (L) NIH 3T3 cells were transfected with ITGA11 ‐overexpression vector or GFP and subjected to western blot analysis to detect the ITGA11 and α‐SMA expression levels (see Section ). (M) Migration toward fibronectin (20 µg·mL −1 ) was assessed with ITGA11 ‐overexpressing NIH 3T3 cells. (M; n = 8, Mann–Whitney) Vertical axis: number of migrated cells per 5HPF. The values represent the mean ± SD. * P < 0.05, ** P < 0.01, **** P < 0.0001.

Article Snippet: HFL‐1 human fetal lung fibroblasts (catalog no. CCL‐153) and A549 human lung adenocarcinoma (catalog no. CCL‐185) cell lines were purchased from American Type Culture Collection (Manassas, VA, USA); NIH 3T3 murine embryo fibroblast (catalog no. EC93061524‐F0), and BEAS‐2B human normal bronchial epithelium (catalog no. EC95102433) were purchased from European Collection of Authenticated Cell Cultures (Salisbury, UK).

Techniques: Modification, Western Blot, Expressing, Migration, Transfection, Over Expression, Plasmid Preparation, MANN-WHITNEY

Journal: Molecular Therapy Oncolytics

Article Title: Intratumoral delivery of a novel oncolytic adenovirus encoding human antibody against PD-1 elicits enhanced antitumor efficacy

doi: 10.1016/j.omto.2022.04.007

Figure Lengend Snippet: AdC68-spE1A-αPD-1 induced tumor cell death in vitro (A–I) HFL-1 (A), HCT-8 (B), A549 (C), Siha (D), NCI-H508 (E), Huh7 (F), MC38 (G), CT26.WT (H), and YUMM5.2 (I) were infected with adenoviruses at indicated MOIs for 72 h. The viability of different cells was measured by CCK-8 assay. Data are shown as mean ± SEM.

Article Snippet: Human embryonic kidney (HEK) 293 cells, human normal lung fibroblast cell line (HFL-1), human colon adenocarcinoma cell line (HCT-8), human colorectal carcinoma cell line (NCI-H508), human hepatocarcinoma cell line (Huh7), and murine melanoma cell line (YUMM5.2) were purchased from ATCC (Manassas, VA, USA).

Techniques: In Vitro, Infection, CCK-8 Assay

Journal: Molecular Therapy Oncolytics

Article Title: Intratumoral delivery of a novel oncolytic adenovirus encoding human antibody against PD-1 elicits enhanced antitumor efficacy

doi: 10.1016/j.omto.2022.04.007

Figure Lengend Snippet: AdC68-spE1-αPD-1 induced tumor cell death by activating apoptotic and autophagic pathways NCI-H508 and Huh7 cells were infected with the indicated adenoviruses at 20 MOIs. The cells were collected at 24 and 48 hpi to conduct the western blot assay under denaturing condition. The proteins p62, LC3 Ι / Π , cleaved caspase-3, cleaved PARP, p53, p21, Mcl-1, CyclinD1, and CyclinE1 were analyzed. Actin was used as a loading control.

Article Snippet: Human embryonic kidney (HEK) 293 cells, human normal lung fibroblast cell line (HFL-1), human colon adenocarcinoma cell line (HCT-8), human colorectal carcinoma cell line (NCI-H508), human hepatocarcinoma cell line (Huh7), and murine melanoma cell line (YUMM5.2) were purchased from ATCC (Manassas, VA, USA).

Techniques: Infection, Western Blot