Fig 2: Tanreqing injection (TRQ) inhibits inflammatory responses in LPS-activated macrophages. (A–D) The toxicity of TRQ at different concentrations in RAW264.7 cells, as determined by CCK-8 assay. (E) Apoptosis assays of RAW264.7 cells treated with TRQ at concentrations of 1:64, 1:128 and 1:256. (F) The expression of SNHG1 in LPS-activated macrophages after TRQ treatment, as determined by qRT-PCR. (G) Expression of proinflammatory cytokines (TNF-a, iNOS, IL-18, MCP-1, IL-6 and IL-1ß) in LPS-activated RAW264.7 cells after TRQ treatment, as determined by qRT-PCR. (H, I) Expression of proinflammatory cytokines (TNF-a, IL-6) in LPS-activated RAW264.7 cells after TRQ treatment, as determined by ELISA. (J) Production of ROS in RAW264.7 cells after LPS and TRQ treatment (magnification, ×100, scale bar, 100 µm). (K, L) Relative protein levels of p-NF-?B-p65, NF-?B-p65, p-MAPK-p38, MAPK-p38 and HMGB1 were detected in LPS-induced RAW264.7 cells treated with TRQ. Data are indicated as the mean ± SD, ns P=0.05, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Fig 3: AuNP@mSiO2@DOX-ZnO induced ICD by promoting tumor cells to release DAMP and DC maturation in vitro and in vivo. (A) Flow cytometric analysis of the CRT exposure on the surface of B16/F10 melanoma cells after treatment with PBS, AuNP@mSiO2, AuNP@mSiO2-ZnO or AuNP@mSiO2@DOX-ZnO with or without irradiation by 655 nm laser at 1.0 W/cm2 for 5 min as indicated. The red line indicates Alexa Fluor 488-CRT, and the black line indicates isotype. (B, D) HMGB1 and ATP secretion in culture supernatants after treatment for 72 h and 48 h, respectively. (C) The expression of intracellular HMGB1 was detected after B16/F10 melanoma cells were treated as indicated for 24 h and 48 h. (E, F) The percentages of CD11c+DCs and CD11c+CD80+CD86+DCs in TDLNs of tumor-bearing mice after treatment for 72 h were identified by flow cytometric analysis (n = 3). L refers to laser irradiation. Results are representative of at least three independent experiments. Statistical significance was calculated by the t-test. Data was presented as mean ± SD. The error bars represent the standard deviation. *P < 0.05, **P < 0.01. ns refers to not significant. TDLNs: tumor draining lymph nodes.

Fig 4: SNHG1 interacts with HMGB1 in RAW264.7 cells. (A) The results of the silver staining assay showed the SNHG1 binding protein. (B) GO function analysis of differentially expressed proteins that bind SNHG1. (C) KEGG pathway analysis of differentially expressed proteins that bind SNHG1. (D) Two HMGB1 peptides were detected in the mass spectrometry results. (E, F) The expression of HMGB1 in RAW264.7 cells treated with LPS, as determined by Western blot. (G) Anti-HMGB1 RIP assay was executed in RAW264.7 cells, followed by qRT-PCR. (H) Expression of proinflammatory cytokines (TNF-a, iNOS, IL-18, MCP-1, IL-6 and IL-1ß) in RAW264.7 cells transfected with lentivirus sh-HMGB1 treated with LPS, as determined by qRT-PCR. (I, J) Expression of proinflammatory cytokines (TNF-a, IL-6) in RAW264.7 cells transfected with lentivirus sh-HMGB1 treated with LPS, as determined by ELISA. Data are indicated as the mean ± SD, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Fig 5: TRQ mitigates LPS-induced ALI. (A) Ultrasound imaging was performed to assess lung injury in the control (a), LPS (b), LTL (c), LTM(d), LTH (e) and TRQ (f) groups. (B) HE staining was performed to assess lung injury in the control (a), LPS (b), LTL (c), LTM (d), LTH (e) and TRQ (f) groups (magnification, ×200, scale bar, 50 µm). (C) The lung injury score of HE staining. (D) The TUNEL apoptosis assay of lung tissue in the control (a), LPS (b), LTL (c), LTM (d), LTH (e) and TRQ (f) groups (magnification, ×100, scale bar, 100 µm). (E) The apoptosis cells number of lung tissue. (F) The wet/dry weight ratio of right upper lung tissue in the control, LPS, LTL, LTM, LTH and TRQ groups. (G–I) Expression of proinflammatory cytokines (TNF-a, IL-6 and HMGB1) in the BALF, as determined by ELISA. (J–L) Expression of proinflammatory cytokines (TNF-a, IL-6 and HMGB1) in the peripheral blood supernatant, as determined by ELISA. Data are indicated as the mean ± SD, ns, P =0.05, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.