Alexa fluor 488 conjugated goat anti rabbit igg h l

Bamboo-leaf flavonoids (the mass fraction of four carbon glycoside flavonoids was 80.66%, Supplementary Figure S2) were prepared in laboratory. L-α-Phosphatidylcholine, cholesterin, CH, AL, 2–2′azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), 2,2-Diphenyl-1-picrylhydrazyl (DPPH), 2,2′-Azobis (2-methylpropionamidine) dihydrochloride (AAPH), 3-(4,5-dimthyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT), 4′,6-diamidino-2-phenylindole (DAPI) were obtained from Sigma Aldrich (St. Louis, MO, USA). Reagents required for cell culture were purchased from HyClone (Logan, UT, USA) and Gibco (Grand Island, NY, USA), respectively. K9M-H3 and Alexa Fluor 488-conjugated Goat Anti-Rabbit IgG (H + L) antibodies were purchased from ABclonal Technology (Wuhan, Hubei, China); p21, p16 and Goat Anti-Rabbit secondary antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA). HPLC-grade methanol was obtained from TEDIA (Cincinnati, OH, USA). The 10 kDa dialysis bag was provided by Spectrum (San Jose, CA, USA). Other chemicals and reagents were purchased from Sinopharm (Shanghai, China).

The cells (A549 and H1299) were seeded onto coverslips in 24‐well plates and cultured at 37°C incubator for 24 h, after cells were adhered on coverslips, taken out and fixed with 4% paraformaldehyde for 30 min, PBS wash 3 times, then permeabilized by fresh 0.5% Triton X‐100 and blocked with 1% bovine serum albumin (BSA). Immunofluorescence analysis assay was performed as described previously.³¹ Primary antibodies against SELENBP1 (1:200 dilution, Abcam, ab90135) and GPX1 (1:100 dilution, ABclonal, A1110), overnight at 4°C, followed by the detection with Alexa Fluor 488‐conjugated Goat Anti‐Rabbit IgG (H + L; 1:100 dilution, ABclonal, AS053) or Alexa Fluor 594‐conjugated Goat Anti‐Rabbit IgG (H + L; 1:100 dilution, ABclonal, AS039). Pictures were taken under a Zeiss Imager Z2 microscope (Carl Zeiss; 400×).

BLF (the mass fraction of orientin, isoorientin, vitexin, and isovitexin was 80.66%, Figure S2) were prepared in the laboratory. AAPH, rapamycin (Rapa), and 3-methyladenine (3-MA) were obtained from Sigma (St.Louis, MO, USA). Dehydrocorydaline (DE) and SB203580 (SB) were obtained from MedChemExpress (Deer Park, NJ, USA). Reagents required for cell culture were purchased from HyClone (Logan, UT, USA) and Gibco (Grand Island, NY, USA), respectively. Anti-JNK1/2/3 and p-JNK1/2/3 (Thr183/Thr183/Thr221) antibodies were purchased from Beyotime biotechnology (Shanghai, China); anti-K9M-H3, IL-10, and Alexa fluor 488-conjugated goat anti-rabbit IgG (H+L) antibodies were purchased from ABclonal technology (Wuhan, China); anti-p16, p21, COX-2, LC3A/B, Beclin-1, SQSTM1/p62, p38, p-p38 (Thr180/Tyr182), ERK1/2, p-ERK1/2 (Thr202/Tyr204), ULK1, MMP-3, COL1A1, GAPDH, and anti-rabbit IgG, HRP-linked antibodies were obtained from Cell signaling technology (Beverly, MA, USA).