human csf2 elisa kit  (Proteintech)

Journal: Journal of translational medicine

Article Title: CircPRKCA facilitates esophageal squamous cell carcinoma metastasis via m5C-dependent CSF2 mRNA stabilization.

doi: 10.1186/s12967-025-06395-5

Figure Lengend Snippet: Fig. 6 CSF2 is a downstream gene of circPRKCA in TE1 and KYSE30 cells. A Differentially expressed genes in TE1 cells transfected with si-circPRKCA were identified using mRNA sequencing. B CSF2 was identified in both the circPRKCA mRNA sequencing dataset and the YBX1-related gene dataset from the TCGA database. C, D The expression of CSF2 in ESCC cells treated with si-NC/si-circPRKCA and pLC5/circPRKCA were measureed by RT-qPCR. E–G Rescue experiments confirmed the changes in scratch healing, transwell migration, and matrigel invasion capabilities of KYSE150 cells under conditions of exogenously added rhCSF2 or the knockdown of circPRKCA, as well as the angiogenic potential of HUVEC cells cultured with medium from KYSE150 cells. H, I RT-qPCR and ELISA were employed to assess the expression of CSF2 in ESCC cells modulated with circPRKCA or si-YBX1. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Cell culture supernatants were collected and analyzed for cytokine production using a Human CSF2 ELISA kit (Proteintech, Wuhan, China) according to the manufacturer’s protocol.

Techniques: Transfection, Sequencing, Expressing, Quantitative RT-PCR, Migration, Knockdown, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Journal of translational medicine

Article Title: CircPRKCA facilitates esophageal squamous cell carcinoma metastasis via m5C-dependent CSF2 mRNA stabilization.

doi: 10.1186/s12967-025-06395-5

Figure Lengend Snippet: Fig. 7 CircPRKCA/YBX1 enhances the stability of CSF2 mRNA through m5C modification in KYSE30 and KYSE150 cells. A RIP assay verifies the interaction between YBX1 and CSF2. B MeRIP assay confirms the presence of m5C methylation sites in CSF2. C, D Actinomycin D experiments assess the regulation of CSF2 mRNA stability by YBX1. E, F A MeRIP experiment verifies the changes in CSF2 mRNA enrichment levels induced by the m5C antibody in ESCC cells transfected with circPRKCA or si-YBX1. G, H Salvage experiments confirm the effect of actinomycin D on the stabilization of CSF2 mRNA in ESCC cells regulated by circPRKCA or si-YBX1. I, J. The levels of YBX1, CSF2, CD31, and CD34 in the pLC5 and circPRKCA vector groups were detected by IHC. K The mechanism of the circPRKCA/YBX1/CSF2 axis in promoting the progression of ESCC metastasis. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: Cell culture supernatants were collected and analyzed for cytokine production using a Human CSF2 ELISA kit (Proteintech, Wuhan, China) according to the manufacturer’s protocol.

Techniques: Modification, Methylation, Transfection, Plasmid Preparation

Journal: Cell death & disease

Article Title: Sanshool improves UVB-induced skin photodamage by targeting JAK2/STAT3-dependent autophagy.

doi: 10.1038/s41419-018-1261-y

Figure Lengend Snippet: Fig. 3 Sanshool treatment induces autophagy and the CSF2-dependent JAK2–STAT3 signaling pathway in UVB-irradiated HDFs. a HDFs treated with sanshool for 24 h after UVB irradiation; analysis of CSF2 secretion by ELISA; UVB-irradiated HDF or non-irradiated HDFs treated with sanshool when co-cultured with CSF2 (0.1 ng/mL). b Expression levels of LC3I/II, p62, MMP-1, MMP-3, and β-actin measured by Western blot analysis. c Representative images of LC3 staining by measuring fluorescence intensity in HDFs in different groups of cells infected with the RFP-GFP-LC3 adenovirus for 24 h by fluorescence microscopy. d Quantitation of punctate structures per cell, based on the number of punctate structures in GFP- RFP-LC3-positive cells. Addition of a CSF2-neutralizing antibody (5 μg/mL) to the medium in each group and incubation with sanshool in UVB- irradiated HDFs; e Expression levels of LC3I/II, p62, MMP-1, MMP-3, and actin measured by Western blot analysis; f Cell viability in response to CSF2 depletion or CSF2 oversupply. g Expression levels of JAK2, phospho-JAK2, STAT3, phospho-STAT3, AKT, and phospho-AKT after sanshool and/or AZD1480 treatment of UVB-irradiated or non-irradiated HDFs for 24 h, determined by Western blot analysis. (h) Relative protein expression levels of AKT and phospho-AKT. i Relative protein expression levels of JAK2 and phospho-JAK2; j Relative protein expression levels of STAT3 and phospho- STAT3. Results presented as means ± SD of three independent experiments (n = 3). *Compared with the UVB-treated group, P < 0.05

Article Snippet: ELISA and Western blot assays Secretion of MMP-1(P03956), MMP-3 (P08254, Raybiotech, Inc.), and CSF2 (DGM00, R&D Systems) was detected using an ELISA kit in accordance with the instruction provided by the manufacturer.

Techniques: Irradiation, Enzyme-linked Immunosorbent Assay, Cell Culture, Expressing, Western Blot, Staining, Infection, Microscopy, Quantitation Assay, Incubation