Journal: International Journal of Molecular Sciences

Article Title: Synovial CXCL3 + FOSL2 + Macrophages Mediate Inflammation via FOSL2 /AP-1 in Rheumatoid Arthritis: A Single-Cell Transcriptome Analysis

doi: 10.3390/ijms26199718

Figure Lengend Snippet: M1/M2 polarization in vitro after knockdown of FOSL2 . ( A ) The workflow of M1/M2 polarization in vitro using U937 cells infected by shRNA- FOSL2 -containing lentivirus; ( B ) The comparisons of M1 markers after knockdown of FOSL2 ; Statistical significance was determined by one-way ANOVA, *, p < 0.05; **, p < 0.01; ***, p < 0.001; ns, non-significant. ( C ) The gene set enrichment among shRNA (M0/M1/M2) and control groups, with the red box highlighting the M1 cells without FOSL2 knockdown as the control.

Article Snippet: Briefly, lentiviral vectors carrying shRNA targeting FOSL2 (targeted sequencing, 5′-GCAGTGAGTATTGGAAGACTT-3′) and a non-specific shRNA control (target sequencing, 5′-CCTAAGGTTAAGTCGCCCTCG-3′) were constructed and provided by OBiO Technology (Shanghai, China).

Techniques: In Vitro, Knockdown, Infection, shRNA, Control

Journal: bioRxiv

Article Title: Phospholipase C β4 promotes RANKL-dependent osteoclastogenesis by interacting with MKK3 and p38 MAPK

doi: 10.1101/2024.03.19.585823

Figure Lengend Snippet: PLCβ4 knockdown reduces RANKL-mediated osteoclast differentiation. BMMs were transduced with non-specific control shRNA (Con-sh) or PLCβ4 shRNA (PLCβ4-sh) lentiviral particles. a The expression of PLCβ4 was analyzed by real-time PCR in BMMs (day 0) or preosteoclasts (day 2). b , c , d Transduced BMMs were cultured with M-CSF (30 ng/mL) and the indicated doses of RANKL for 4 or 5 days. b Osteoclasts were visualized after tartrate-resistant acid phosphatase (TRAP) staining and, c the number of osteoclasts was counted. d TRAP activity was determined by using the cultured cell supernatant generated in ( b) . e Transduced BMMs were cultured in the presence of various concentrations of M-CSF. After 3 days, the MTS assay was performed, as described in the Materials and Methods section. f Transduced BMMs were cultured in osteoclastogenic media for the indicated times. Real-time PCR was performed to assess the gene expression of osteoclastogenic markers. All data are expressed as the mean ± SD. * P < 0.05 and ** P < 0.001 vs. the control (Con-sh)

Article Snippet: The lentiviral constructs containing PLCβ4 small hairpin RNA (shRNA) or a non-specific shRNA control (Con-sh) were obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Transduction, shRNA, Expressing, Real-time Polymerase Chain Reaction, Cell Culture, Staining, Activity Assay, Generated, MTS Assay